Search Results (104)

The soil-borne fungal pathogen Verticillium dahliae causes devastating vascular wilt disease in numerous crops, including cotton. In this study, we reveal that VdSOX1, a highly conserved sarcosine oxidase gene, is significantly upregulated during host infection and plays a multifaceted role in fungal physiology and pathogenicity. Functional deletion of VdSOX1 leads to increased fungal virulence, accompanied by enhanced microsclerotia formation, elevated carbon source utilization, and pronounced upregulation of effector genes, including over 50 predicted secreted proteins genes. Moreover, the VdSOX1 knockout strains suppress the expression of key defense-related transcription factors in cotton, such as WRKY, MYB, AP2/ERF, and GRAS families, thereby impairing host immune responses. Transcriptomic analyses confirm that VdSOX1 orchestrates a broad metabolic reprogramming that links nutrient acquisition to immune evasion. Our findings identify VdSOX1 as a central regulator that promotes V. dahliae virulence by upregulating effector gene expression and suppressing host immune responses, offering novel insights into the molecular basis of host–pathogen interactions and highlighting potential targets for disease management. Full article

Microbe–microbe interactions have been explored at the molecular level to a lesser degree than plant–pathogen interactions, primarily due to the economic impact of crop losses caused by pathogenic microorganisms. Effector proteins are well known for their role in disease development in many plant–pathogen pleinteractions, but there is increasing evidence showing their involvement in other types of interaction, including microbe–microbe interactions. Through the use of LC-MS/MS sequencing, effector candidates were identified in the in vitro interaction between a banana pathogen, Pseudocercospora fijiensis and a biological control agent, Trichoderma harzianum. The diverse interaction secretome revealed various glycoside hydrolase families, proteases and oxidoreductases. T. harzianum secreted more proteins in the microbial interaction compared to P. fijiensis, but its presence induced the secretion of more P. fijiensis proteins that were exclusive to the interaction secretome. The interaction secretome, containing 256 proteins, was screened for effector candidates using the algorithms EffHunter and WideEffHunter. Candidates with common fungal effector motifs and domains such as LysM, Cerato-platanin, NPP1 and CFEM, among others, were identified. Homologs of true effectors and virulence factors were found in the interaction secretome of T. harzianum and P. fijiensis. Further characterization revealed a potential novel effector of T. harzianum. Full article

The yellow peach moth (YPM), Conogethes punctiferalis, is an important agricultural insect pest causing severe damage to corn in eastern China. Beauveria bassiana is an effective, eco-friendly, and promising alternative agent for controlling this insect pest. However, insect immunity can limit the ability of fungal infections. In order to understand the immune response mechanism of YPM, a comparative proteomic analysis of non-infected and B. bassiana-infected larvae was conducted using the isobaric tags for relative and absolute quantification (iTRAQ) technique. On the basis of proteomic analysis, 4195 quantifiable proteins were identified from a total of 29,155 peptides. The functions of the identified proteins were annotated in four databases (GO, COG, KEGG, and IPR). A total of 132 immune-related proteins were screened, including 46 pathogen recognition proteins, 27 extracellular signal modulation proteins, and 59 immune effectors. Furthermore, 70 differentially expressed proteins (DEPs) were identified, including 57 up-regulated proteins and 13 down-regulated proteins. Among these, four DEPs were related to immunity, namely one defense protein and three peptidoglycan recognition proteins. Six randomly selected immune-related proteins associated with target genes were validated for qRT-PCR, and the results indicated that the accuracy and reliability of the proteome sequencing data were high. Taken together, the results enrich the fundamental knowledge of YPM immune responses to B. bassiana infection and provide a new insight into insect−pathogen interactions. Full article

Penicillium expansum, a major postharvest pathogen, causes blue mold decay in apples, resulting in substantial economic losses and mycotoxin contamination. Despite the importance of effector proteins in fungal pathogenicity, the role of metalloproteases in P. expansum remains unclear. Here, we characterize an effector candidate, PeMep, through whole genome sequencing and functional analyses. Functional validation confirmed the secretory capacity of its signal peptide via yeast assays and subcellular localization. Deletion of PeMep significantly impaired fungal growth (23% reduction), conidiation (23.3% decrease), and germination efficiency. The ΔPeMep mutant exhibited hypersensitivity to osmotic, oxidative, and thermal stresses, highlighting its vital role in environmental adaptability. Importantly, pathogenicity assays revealed attenuated virulence in the ΔPeMep mutant, with 15–30% smaller lesion sizes on apples and delayed hyphal penetration compared to the wild-type, demonstrating that PeMep is essential for the pathogenic process of P. expansum 3.3703. These findings identify PeMep as a potential multifunctional effector protein crucial for fungal development, environmental adaptation, and pathogenicity in P. expansum 3.3703, providing a novel target for postharvest disease management. Full article

Fungal phytopathogens employ effector proteins and secondary metabolites to subvert host immunity. Effector proteins have attracted widespread interest in infection, especially for unknown, unreported genes. However, the type of protein remains much less explored. Here, we combined transcriptome analysis and functional validation to identify virulence-associated genes in Fusarium graminearum during fungi infection. A unique secreted protein, FGSE02, was significantly upregulated in the early infection stage. Proteomic characterization revealed that the protein contains a functional signal peptide but lacks known domains. The transient expression of FGSE02 in Nicotiana benthamiana induced rapid cell death, while gene knockout stains reduced fungal virulence without affecting growth. Our findings highlight FGSE02 as a key virulence factor, offering potential targets for disease control. Taken together, the results of this study identify a pathogenic factor and provide new insights into the development of green pesticides. Full article

Common in Fungal Extracellular Membrane (CFEM) effectors, a unique class of fungal-specific proteins, play critical roles in host-pathogen interactions. While CFEM proteins have been extensively characterized in phytopathogens, their presence and functions in Fusarium verticillioides remained unexplored. Here, we systematically identified 19 CFEM-containing proteins in F. verticillioides, among which FvCFEM12 exhibited secretory activity and plant infection-induced expression. Functional characterization revealed that FvCFEM12 suppressed Bax- and INF1-triggered cell death in Nicotiana benthamiana leaves. Furthermore, heterologous expression of FvCFEM12 in maize leaves using P. syringae strain D36E can compromise immune responses against bacterial pathogens. Deletion of FvCFEM12 impaired fungal virulence, altered hyphal morphology, and reduced cell wall stress tolerance. Interestingly, FvCFEM12 physically interacted with the maize wall-associated receptor kinase ZmWAK17ET, and targeted silencing of ZmWAK17 in maize enhanced susceptibility to F. verticillioides. Our findings revealed that FvCFEM12 is a dual-function effector that suppresses plant immunity and maintains fungal cell wall integrity, thereby orchestrating fungal pathogenicity at the host–pathogen interface. Full article

Cross-kingdom infections, where pathogens from one kingdom infect organisms of another, were historically regarded as rare anomalies with minimal concern. However, emerging evidence reveals their increasing prevalence and potential to disrupt the delicate balance between plant, animal, and human health systems. Traditionally recognized as plant-specific, a subset of phytopathogens, including certain fungi, bacteria, viruses, and nematodes, have demonstrated the capacity to infect non-plant hosts, particularly immunocompromised individuals. These pathogens exploit conserved molecular mechanisms, such as immune evasion strategies, stress responses, and effector proteins, to breach host-specific barriers and establish infections. Specifically, fungal pathogens like Fusarium spp. and Colletotrichum spp. employ toxin-mediated cytotoxicity and cell-wall-degrading enzymes, while bacterial pathogens, such as Pseudomonas syringae, utilize type III secretion systems to manipulate host immune responses. Viral and nematode phytopathogens also exhibit molecular mimicry and host-derived RNA silencing suppressors to facilitate infections beyond plant hosts. This review features emerging cases of phytopathogen-driven animal and human infections and dissects the key molecular and ecological determinants that facilitate such cross-kingdom transmission. It also highlights critical drivers, including pathogen plasticity, horizontal gene transfer, and the convergence of environmental and anthropogenic stressors that breach traditional host boundaries. Furthermore, this review focuses on the underlying molecular mechanisms that enable host adaptation and the evolutionary pressures shaping these transitions. To address the complex threats posed by cross-kingdom phytopathogens, a comprehensive One Health approach that bridges plant, animal, and human health strategies is advocated. Integrating molecular surveillance, pathogen genomics, AI-powered predictive modeling, and global biosecurity initiatives is essential to detect, monitor, and mitigate cross-kingdom infections. This interdisciplinary approach not only enhances our preparedness for emerging zoonoses and phytopathogen spillovers but also strengthens ecological resilience and public health security in an era of increasing biological convergence. Full article

Fusarium wilt caused by Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) is the most destructive disease of the banana. Effectors play a crucial role in Foc TR4–banana interaction; however, only a few effectors have been functionally characterized. Our previous secretome studies on Foc TR4 highlighted an uncharacterized protein without any conserved domains (named FoUpe9), which was predicted to be a candidate effector. Herein, bioinformatics analysis showed that FoUpe9 was highly conserved among Fusarium species. FoUpe9 was highly induced during the early infection stages in the banana. A yeast signal sequence trap assay showed that FoUpe9 is a secretory protein. FoUpe9 could inhibit cell death and ROS accumulation triggered by BAX through the Agrobacterium-mediated Nicotiana benthamiana expression system. Subcellular location showed that FoUpe9 was located in the nucleus and cytoplasm of N. benthamiana cells. Deletion of the FoUpe9 gene did not affect mycelial growth, conidiation, sensitivity to cell-wall integrity, or osmotic and oxidative stress, but significantly attenuated fungal virulence. FoUpe9 deletion diminished fungal colonization and induced ROS production and expression of SA-related defense genes in banana plants. These results suggest that FoUpe9 enhances Foc TR4 virulence by inhibiting host immune responses and provide new insights into the functions of the uncharacterized proteins, further enhancing our understanding of effector-mediated Foc TR4 pathogenesis. Full article

Puccinia polysora Underw, the pathogen that causes southern corn rust (SCR), delivers effectors to manipulate host immune responses. However, the mechanisms by which these effectors modulate host defenses are not well characterized. In this study, we found that the P. polysora effector PpEX is highly upregulated during infection. PpEX suppresses plant immune responses that are initiated by chitin, including the activation of mitogen-activated protein kinases (MAPKs) and the expression of pathogenesis-related (PR) genes. Maize plants transiently expressing PpEX exhibited higher pathogen infection rates, larger colony areas, and greater fungal biomass on their leaves compared to the control group. By employing TurboID proximity labeling technology coupled with mass spectrometry analysis, we discovered potential target proteins of PpEX in maize. The split-luciferase system enabled us to identify ZmMPK3, a component of the MAPK signaling pathway, as an interacting partner of PpEX among the candidate proteins. This interaction was subsequently confirmed by co-immunoprecipitation (Co-IP) experiments. Additionally, we verified that ZmMPK3 plays a positive role in regulating maize resistance to SCR. Thus, PpEX may function as a virulence effector that dampens plant PTI immunity by interacting with ZmMPK3 and impeding the MAPK signaling pathway. Full article

Plants are challenged regularly with multiple types of biotic stress factors, such as pathogens or insect herbivores, in their environment. To detect and defend against pathogens, plants have evolved an innate immune system in which intracellular receptors in the so-called effector-triggered immunity play a vital role. In Arabidopsis thaliana the Toll/interleukin-1 receptors (TIRs) domain is related to intracellular immunity receptors, for example in TIR-NBS-LRR (TNL) proteins. Among the TIR domain carrying proteins, very little is known about the function of the TIR-X proteins. Here, we focus on the recently described TIR-X (TIRP; At5g44900) to analyze its role in phytohormone-mediated plant defense through gene expression and phytohormone quantification. Therefore, we employed two fungal pathogens, the necrotrophic Alternaria brassicicola and the hemibiotrophic Verticillium dahliae, to infect A. thaliana WT (Col-0), TIRP knock-out, and TIRP overexpressing lines for comparative analyses. Furthermore, we included the insect herbivore Spodoptera littoralis and a treatment with S. littoralis egg extract on the plants to analyze any role of TIRP during these attacks. We found that both A. brassicicola and V. dahliae infections increased TIRP gene expression systemically. The salicylic acid content was higher in the TIRP overexpressing line, corresponding to a better S. littoralis larval growth performance in feeding assays. However, since we never observed clear infection-related differences in jasmonate or salicylic acid levels between the wild type and the two transgenic Arabidopsis lines, our results rule out the possibility that TIRP acts via the regulation of phytohormone synthesis and accumulation. Full article

The biotrophic fungus Ustilago maydis, which causes smut disease in maize, secretes numerous proteins upon plant colonization. Some of them, termed effectors, help to evade plant defenses and manipulate cellular processes within the host. The function of many proteins specifically secreted during infection remains elusive. In this study, we biochemically characterized one such protein, UMAG_00027, that is highly expressed during plant infection. We show that UMAG_00027 is a secreted protein with a lectin-like fold and therefore term it Llp1 (lectin-like-protein 1). Llp1 decorated the fungal cell wall of cells grown in axenic culture or proliferating in planta, which is in agreement with its potential sugar-binding ability. We were unable to identify the precise sugar moieties that are bound by Llp1. CRISPR/Cas9-mediated deletion of llp1 reveals that the gene is not essential for fungal virulence. A structural search shows the presence of several other lectin-like proteins in U. maydis that might compensate for the function of Llp1 in ∆llp1 mutants. We therefore speculate that Llp1 is part of a family of lectin-like proteins with redundant functions. Full article

Bacteriophages, as ubiquitous bacterial viruses in various natural ecosystems, play an important role in maintaining the homeostasis of the natural microbiota. For many years, bacteriophages were not believed to act on eukaryotic cells; however, recent studies have confirmed their ability to affect eukaryotic cells and interact with the host immune system. Due to their complex protein structure, phages can also directly or indirectly modulate immune processes, including innate immunity, by modulating phagocytosis and cytokine reactions, as well as acquired immunity, by producing antibodies and activating effector cells. They can therefore have a profound impact on the course of bacterial infections by stimulating and at the same time inhibiting the systemic pro-inflammatory response. This review article presents a characterization of the processes by which bacteriophages affect selected immune mechanisms in selected animal species. The results of our own experiments using calves are also presented as examples. The paper contains many new examples of potential uses of bacteriophages and their effects on eukaryotic cells, especially in the course of bacterial infections, which are extremely important in experimental treatments exploiting phages as alternatives to antibiotics. The positive results of the effects of bacteriophages on eukaryotic cells during infections open up promising new prospects for their use as natural tools in the treatment of bacterial, fungal, and viral diseases in animals and humans. Full article

Monilinia fructicola is the most common and destructive brown rot agent on peaches. Knowledge of gene expression mediating host–pathogen interaction is essential to manage fungal plant diseases. M. fructicola putative virulence factors have been predicted by genome investigations. The pathogen interaction with the host was validated. Five M. fructicola isolates were inoculated on two cultivars (cv.s) of peach (Prunus persica (L.) Batsch) ‘Royal Summer’ and ‘Messapia’ with intermediate and late ripening periods, respectively. The expression pattern of 17 candidate effector genes of M. fructicola with functions linked to host invasion and fungal life, and seven peach genes involved in the immune defense system were monitored at 0, 2, 6, 10, and 24 h-post inoculation (hpi). All fungal isolates induced similar brown rot lesions on both cv.s whereas the modulation of effector genes was regulated mainly at 2, 6, and 10 hpi, when disease symptoms appeared on the fruit surface, confirming the involvement of effector genes in the early infection stage. Although differences were observed among the fungal isolates, the principal component investigation identified the main differences linked to the host genotype. The salicylic acid and jasmonate/ethylene signaling pathways were differently modulated in the host independent from the fungal isolate used for inoculation. On plants susceptible to brown rot, the pathogen may have adapted to the host’s physiology by modulating its effectors as weapons. Full article

Rehmannia glutinosa is an important medicinal herb; but its long-term cultivation often leads to continuous cropping problems. The underlying cause can be attributed to the accumulation of and alterations in root exudates; which interact with soil-borne pathogens; particularly Fusarium oxysporum; triggering disease outbreaks that severely affect its yield and quality. It is therefore crucial to elucidate the mechanisms by which root exudates induce F. oxysporum CCS043 outbreaks. In this study; the genome of F. oxysporum CCS043 from R. glutinosa’s rhizosphere microbiota was sequenced and assembled de novo; resulting in a 47.67 Mb genome comprising 16,423 protein-coding genes. Evolutionary analysis suggests that different F. oxysporum strains may adapt to the host rhizosphere microecosystem by acquiring varying numbers of specific genes while maintaining a constant number of core genes.The allelopathic effects of ferulic acid; verbascoside; and catalpol on F. oxysporum CCS043 were examined at the physiological and transcriptomic levels. The application of ferulic acid was observed to primarily facilitate the proliferation and growth of F. oxysporum CCS043; whereas verbascoside notably enhanced the biosynthesis of infection-related enzymes such as pectinase and cellulase. Catalpol demonstrated a moderate level of allelopathic effects in comparison to the other two. Furthermore; 10 effectors were identified by combining the genomic data. Meanwhile; it was found that among the effector-protein-coding genes; ChiC; VRDA; csn; and chitinase exhibited upregulated expression across all treatments. The expression patterns of these key genes were validated using qRT-PCR. Transient overexpression of the two effector-encoding genes in detached R. glutinosa leaves provided further confirmation that ChiC (GME8876_g) and csn (GME9251_g) are key effector proteins responsible for the induction of hypersensitive reactions in R. glutinosa leaf cells. This study provides a preliminary indication that the use of allelochemicals by F. oxysporum CCS043 can promote its own growth and proliferation and enhance infection activity. This finding offers a solid theoretical basis and data support for elucidating the fundamental causes of fungal disease outbreaks in continuous cropping of R. glutinosa and for formulating effective mitigation strategies. Full article

Helotiales, a diverse fungal order within Leotiomycetes (Ascomycota), comprises over 6000 species occupying varied ecological niches, from plant pathogens to saprobes and symbionts. Despite their importance, their genetic adaptations to temperature and environmental conditions are understudied. This study investigates temperature adaptations in infection genes and substrate degradation genes through a comparative genomics analysis of 129 Helotiales species, using the newly sequenced genomes of Gyoerffyella rotula and Anguillospora crassa. Key gene families such as cytochrome P450 enzymes, virulence factors, effector proteins, and carbohydrate-active enzymes (CAZymes) were analyzed to understand their roles in temperature and lifestyle adaptations, uncovering possible alternative lifestyle mechanisms. Our findings reveal that Helotiales fungi possess genes associated with nutrient acquisition, pathogenicity, and symbiotic relationships strongly adapted to cold environments that might be impacted by global warming. On the other hand, some species demonstrate potential for adaptation to warmer climates, suggesting increased activity in response to global warming. This study reveals the adaptive mechanisms enabling Helotiales fungi to thrive in both cold and warm environments. These findings provide valuable insights into their ecological success and evolutionary resilience, which may facilitate their ability to transition between pathogenic, symbiotic, and saprobic phases in response to changing environmental conditions. Full article