Global wheat production faces growing threats from rising temperature and pathogen attacks under climate change. Here, we developed a nanoenabled seed priming strategy that simultaneously enhanced wheat thermotolerance and resistance to Fusarium head blight (FHB). Specifically, under heat shock stress (45 °C, 24 h), wheat seedlings that underwent AgSiO2NP (40 mg/L AgNPs + 40 mg/L SiO2NPs) seed priming exhibited reduced wilting symptoms, increased biomass (22.8%), improved water uptake, maintained cellular membrane integrity, and delayed protein/starch catabolism compared to the hydroprimed control. Reactive oxygen species (ROS) localization assays and RNA-seq analysis reveal that AgSiO2NP priming triggered ROS-mediated activation of "plant hormone signal transduction" and the "MAPK signaling pathway" in seeds. These molecular changes persisted into the seedling stage, establishing a primed state that enhanced seedlings' heat resilience. AgSiO2NP-primed seedlings also showed enhanced resistance to Fusarium graminearum, even under hotter conditions (28 °C). Comparative transcriptomics of wheat seedlings pre- and postpathogen infection revealed that AgSiO2NP priming intensified defense-related pathways, including the MAPK signaling pathway, plant hormone signal transduction pathway, plant-pathogen interaction pathway, and specialized metabolite biosynthesis pathways, as compared to hydropriming, leading to robust immune activation and disease resistance during pathogen attack. We also demonstrate that an inexpensive ($0.5-2 per acre) and environmentally friendly alternative, CuO@SiO2 nanoparticles, efficiently enhances the resistance of wheat seedlings to FHB. This study proposes a nontransgenic approach for engineering climate-resilient wheat, providing a sustainable strategy to address food insecurity exacerbated by climate change.

Liquid-liquid phase separation (LLPS) has emerged as an important strategy for plant stress resistance, yet its dynamic regulation during plant-pathogen interaction remains poorly understood. Here, we demonstrate that the stripe rust fungus Puccinia striiformis f. sp. tritici (Pst) deploys effector Hasp170 (one haustorial secreted protein) to subvert wheat immunity by directly disrupting host LLPS. Hasp170 targets the intrinsically disordered region (IDR1) of the wheat nuclear protein TaPSTE (phase separation protein targeted by effector), which forms LLPS-dependent biomolecular condensates. Within these condensates, TaPSTE recruits the transcription factor TaNF-YC, thereby activating the expression of genes driving reactive oxygen species burst and Ca2+ influx, key components for disease resistance. By binding TaPSTE's IDR1, Hasp170 impairs condensate formation and prevents TaNF-YC recruitment, consequently suppressing host immunity and facilitating fungal parasitism. This reveals a virulence strategy where pathogens directly manipulate host biomolecular condensates to evade immune responses.

Spo11-mediated DNA double-strand breaks (DSBs) are essential for meiotic recombination, yet how Spo11 activity is temporally regulated during mitosis and fungal development remains unclear. In the fungal plant pathogen Fusarium graminearum, we found that FgSpo11 has a DSB-independent role delaying meiosis I and a DSB-dependent role critical for postmeiotic mitoses during ascosporogenesis. Loss of FgSpo11 accelerates meiosis I and causes excessive postmeiotic divisions, ultimately causing aborted ascospores. A premature stop codon (TAG) is corrected to tryptophan (TGG) by adenosine-to-inosine RNA editing exclusively during sexual reproduction, enabling full-length protein synthesis. A genomically "corrected" allele bypassing this editing preserves ascospore formation but causes meiotic and vegetative mitotic defects. Beyond its on-switch function, this editing acts as a tunable rheostat fine-tuning FgSpo11 dosage during meiosis. Evolutionary analyses reveal recurrent gain and loss of this editing, highlighting adaptive modulation of Spo11 deployment. This study uncovers a single-site RNA editing gate controlling a key meiotic regulator and illustrates transcriptome plasticity in reconciling life cycle demands in eukaryotic pathogens.

Fusarium graminearum causes Fusarium head blight (FHB), a devastating wheat disease that threatens global food security and safety. Nuclear Dbf2-related (NDR) kinases regulate cellular morphogenesis, but their roles in phytopathogenic fungi remain elusive. Here, we characterized F. graminearum NDR kinase FgCot1, whose deletion severely disrupted vegetative growth, conidiation, polarized growth and pathogenicity. Notably, the Fgcot1 mutant showed genetic instability, producing fast-growing suppressors with primary (Gpmk1 pathway genes), secondary (MFS transporter SSF1) and tertiary (transcription factor TSF1) mutations. Functional analyses revealed that disrupting the Gpmk1 pathway partially rescued the Fgcot1 growth defects due to Gpmk1 hyperactivation, whereas SSF1 deletion independently enhanced growth. The TSF1R386C mutation suppressed the Fgcot1 growth defect via a Gpmk1/Ssf1-independent pathway. Moreover, FST11 deletion and TSF1R386C mutation significantly alleviated the Fgcot1 mutant's polarized growth defect and slightly restored conidiation, suggesting that Gpmk1 and Tsf1 function in parallel pathways downstream of FgCot1 kinase. Finally, we demonstrated that FgCot1 interacts with Tsf1 and regulates its protein stability and nuclear accumulation; R386C mutation restored these defects of Tsf1 in the Fgcot1 mutant. Collectively, FgCot1 kinase integrates Gpmk1 signalling and Tsf1-dependent transcription to govern polarized growth and conidiation in F. graminearum.

Fhb1 is the most widely used locus for Fusarium head blight (FHB) resistance in wheat, yet the mechanistic basis of its candidate gene, TaHRC, remains elusive. Here, we demonstrate that the protein from the resistant allele TaHRC-R localises to both the nucleus and cytoplasm, whereas the susceptible protein TaHRC-S is confined to the nucleus. Remarkably, only TaHRC-R triggered a reactive oxygen species (ROS) burst in planta, dependent on its extranuclear localisation. The N-terminal 21 amino acids that distinguish TaHRC-R from TaHRC-S were essential for its nuclear export and ROS induction. Within the nucleus, TaHRC-R formed heterodimers with TaHRC-S via its N-terminal 21 amino acids and central region, disrupting the formation of large, sparse TaHRC-S condensates and converting them into numerous smaller assemblies. These results reveal a dual, spatially coordinated mechanism whereby TaHRC-R promotes ROS production outside the nucleus while modulating nuclear condensate dynamics to counteract the susceptible allele. This compartmentalised functionality provides a molecular framework for Fhb1-mediated resistance and illustrates a novel paradigm of subcellular specialisation in plant immunity.

Carbon catabolite repression (CCR) acts as a switch, reprogramming nutrient utilization in fungal pathogens during the growth-to-colonization transition. However, whether this regulatory system can be exploited by other microbes remains unknown. Here, we demonstrate that Pseudomonas CXZ-8 attenuates the virulence of Fusarium graminearum by hijacking fungal CCR. CXZ-8 disrupts the infection-induced nuclear-to-cytoplasmic relocalization of the CCR master regulator FgCreA, thereby suppressing FCO1 expression, which is crucial for both host cell wall degradation and nutrient acquisition. This interference also benefits the bacterium by preventing the accumulation of host-derived indole derivatives and fungal mycotoxins that threaten its survival. Notably, approximately 20% of field-isolated bacteria exhibit similar FgCreA-stabilizing activity. Furthermore, we assembled a microbial consortium enriched for CCR-targeting bacteria, which conferred broad-spectrum disease resistance in field trials. These findings reveal a novel mode of interkingdom interference and establish CCR as a conserved microbial vulnerability, with implications for sustainable, microbiome-based crop protection.

Plants reprogramme their metabolism upon pathogen attack, producing compounds that can either enhance immunity or be exploited by pathogens. Metabolomic profiling of wheat during Fusarium graminearum infection revealed pronounced accumulation of phenolamides, driven by activation of their biosynthetic pathways. Notably, exogenous N-feruloylputrescine (Ferput), a representative phenolamide, enhanced resistance to wheat stripe rust, powdery mildew and rice blast but increased susceptibility to Fusarium head blight. Ferput inhibited fungal infection in the lemma while promoting rachis colonisation, indicating pathogen- and tissue-specific effects. Mechanistic analyses showed that Ferput stimulates deoxynivalenol (DON) biosynthesis by inducing TRI gene expression, toxisome formation and DON-associated cellular differentiation, underlying the shift from lemma resistance to rachis susceptibility. Together, these findings highlight the context-dependent roles of phenolamide in plant-pathogen interactions and suggest that, under specific pathological contexts, defence-associated metabolites can be exploited by pathogens to enhance their virulence. This insight underscores the necessity of considering the dual functional roles of plant metabolites when engineering broad-spectrum disease resistance.

AIMS:To isolate and characterize a rhizosphere-derived Bacillus velezensis strain with biocontrol and plant growth-promoting potential, and to evaluate its efficacy against plant pathogens together with its genomic basis. METHODS AND RESULTS:A bacterial strain (BJ-1141) was isolated from the rhizosphere and identified as B. velezensis based on morphological characteristics and phylogenetic analyses of 16S rRNA and gyrA gene sequences. The strain exhibited broad-spectrum antagonistic activity against 24 plant pathogenic fungi, inhibiting 23 pathogens by more than 50%, with a maximum inhibition rate of 95.57%. BJ-1141 produced hydrolytic enzymes (amylase, protease, cellulase, and pectinase) and plant growth-promoting factors, including siderophores and indole-3-acetic acid (IAA). It also demonstrated phosphorus solubilization, nitrogen fixation, and biofilm formation. Greenhouse experiments showed that root application of BJ-1141 effectively controlled tomato Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici, achieving control efficacies of 81.13-85.93%, which were higher than those of chemical and commercial microbial treatments. In addition, BJ-1141 significantly increased plant height, biomass, and chlorophyll content in tomato and lettuce seedlings. Whole-genome sequencing revealed a 3 902 965 bp genome containing 13 biosynthetic gene clusters associated with antimicrobial secondary metabolites, including surfactin, fengycin, bacillaene, difficidin, macrolactin H, bacillibactin, and bacilysin. CONCLUSIONS:Bacillus velezensis BJ-1141 is a multifunctional bacterial strain with strong biocontrol efficacy and plant growth-promoting traits, supported by genomic evidence, and represents a promising candidate for sustainable crop disease management.

Stop codon readthrough is widespread across eukaryotes and often dismissed as translational noise, yet its tissue/stage-specific occurrence suggests adaptive roles in proteome tuning. We asked whether readthrough-related mechanisms can mitigate stage-specific pleiotropic trade-offs without genomic change. In the filamentous ascomycete Fusarium graminearum, the functional solution relies on developmentally programmed A-to-I "stop-loss" RNA editing of the terminal NDR kinase gene FgDBF2 (UAG→UIG, read as UGG), instead of stochastic readthrough. This edit adds a short, intrinsically disordered C-terminal extension acting as a cis-encoded destabilizing element, lowering FgDbf2 dosage during ascospore formation. Genetic and cell biological analyses show meiosis proceeds independently of FgDbf2, but accurate one-nucleus/one-spore encapsulation is promoted by the edited, destabilized isoform. Blocking editing (stop retained) or increasing unedited FgDbf2 yields malformed, multinucleate spores despite normal nuclear counts, establishing ascospore morphogenesis as dosage-sensitive rather than isoform-specific. Conversely, constitutive production of the edited, destabilized isoform impairs vegetative growth and hyphal septation, suggesting stage-specific antagonism with mitotic functions. Mechanistically, the edited tail destabilizes Dbf2 and GFP, likely via nonclassical proteostasis pathways. Epistasis analysis indicates the CDK Cdc2A also restrains FgDbf2 and elevated Cdc2A partially suppresses defects caused by excess unedited FgDbf2. Comparative and transcriptomic analyses reveal conservation of DBF2 stop-loss editing across Sordariomycetes and identify many stop-loss edits encoding destabilizing tails consistent with positive genome-wide selection. We propose that stage-specific stop-loss editing is a developmentally gated dosage-buffering mechanism that transiently reduces NDR kinase abundance during ascospore formation, thereby alleviating growth-reproduction trade-offs without requiring gene duplication.

Maintaining genome integrity is essential, yet how DNA repair is balanced across life stages remains poorly understood. Here we uncover an epitranscriptomic mechanism in the fungal pathogen Fusarium graminearum that alleviates a trade-off between heat-stress adaptation and sexual reproduction. We show that FgMus81 acts independently of its nuclease activity and canonical partner FgMms4, and has dosage-dependent, stage-specific functions: restrained levels support meiosis, whereas elevated levels promote heat-stress survival. We identify a sexual stage-specific A-to-I RNA editing event that recodes FgMus81 (N420D) and tunes its abundance to meet meiotic demands without compromising stress resilience. Notably, both pre-editing and post-editing isoforms support meiotic interhomolog crossovers, but the post-editing isoform impairs mitotic recombination. Conservation of this editing across Sordariomycetes suggests evolutionary selection for stage-specific control. Together, these findings reveal an epitranscriptomic switch that partitions Mus81 functions across life stages and identify adaptive RNA editing as a regulator of homologous recombination in fungal pathogens.

ATP binding is an essential event in diverse biological processes including plant immunity. The ATP-binding domains in plant kinases share similar structural properties, providing a potential common target for pathogens. However, effectors targeting the ATP-binding domains to modulate kinase activity have not been identified. In this study, we identified a conserved effector containing an Egh16-like domain (Cee1) in Fusarium graminearum. As an in planta induced gene, CEE1 plays a stage-specific role in infectious growth within wheat rachis. Upon translocation into plant cells, Cee1 interacts with the ATP-binding domain of TaMPK3 via its Egh16-like domain. This interaction interferes with the ATP binding and impairs the kinase activity of TaMPK3, leading to reduced phosphorylation levels of TaWRKY33 and subsequent inactivation of downstream resistance responses. F. graminearum harbors three paralogs of Cee1, each containing two adjacent motifs responsible for specific interaction with ATP-binding pockets, all crucial for pathogenesis. The quadruple mutant lacking these four CEE genes shows drastically reduced pathogenicity, and CEE genes have been identified as silencing targets for improving wheat FHB resistance. Taken together, Cee1 and its paralogs act as core effectors in F. graminearum by targeting the ATP-binding domains of plant kinases, demonstrating the representative mode of action of the Egh16-like domain in fungal-plant interactions.

Pattern-triggered immunity (PTI) forms the first barrier of plant defense against pathogens, but prolonged immune activation is costly and hampers growth. Therefore, plants employ diverse negative regulators to maintain immune homeostasis. Although intracellular regulators have been well characterized, the roles of extracellular components remain elusive. Notably, plants encode numerous extracellular leucine-rich repeat-only (eLRR) proteins lacking signaling domains, but their functions in immune regulation are largely unexplored. We performed genome-wide screening in Nicotiana benthamiana and identified NbLRRP1, an eLRR protein with high structural similarity to the extracellular domain of central hub NbBAK1 in PTI signaling, suggesting a potential functional association. NbLRRP1 is secreted into the apoplast, where it broadly suppresses PTI induced by multiple pathogen-associated molecular patterns (PAMPs). NbLRRP1-knockout plants display no visible growth defects but markedly enhance resistance to Sclerotinia sclerotiorum. Mechanistically, NbLRRP1 directly interacts with NbBAK1 and disrupts the formation of immune receptor complexes, such as NbBAK1-RE02 and NbBAK1-NbFLS2, thereby attenuating prolonged PTI activation. NbLRRP1 orthologs are broadly distributed among diverse food crops, cash crops, and forest trees, and seem functionally conserved. Collectively, these findings reveal a conserved eLRR-mediated mechanism and highlight this eLRR protein as a promising molecular target for crop disease resistance breeding.

Reactive oxygen species (ROS) produced by respiratory burst oxidase homologs (RBOHs) are critical for plant immunity. Despite transcriptional and post-translational regulation of RBOHD activity, the dynamic control of the ROS burst during plant immune responses remains unclear. Here, we demonstrate that upon infection with avirulent Puccinia striiformis f. sp. tritici (Pst), the wheat transcription factor TaWRKY40 is activated, driving an extracellular ROS burst by binding to the TaNOX10 (RBOHD) promoter and inducing its activation. Furthermore, TaWRKY40 undergoes phosphorylation by the brassinolide signaling kinase TaBSK3, promoting TaWRKY40 nuclear translocation and enhancing TaNOX10 transactivation. This cascade increases extracellular ROS levels, conferring resistance to stripe rust. When encountering virulent Pst races, another wheat WRKY transcription factor, TaWRKY19, is upregulated, transcriptionally suppressing TaWRKY40 and binding to the same cis element in the TaNOX10 promoter. This inhibits TaNOX10 expression, suppresses ROS accumulation, and renders wheat susceptible. These findings reveal a transcriptional activation module comprising TaBSK3-TaWRKY40-TaNOX10 that governs ROS production and establish a TaWRKY19-TaWRKY40 dual-regulatory module that fine-tunes ROS burst during wheat-Pst interactions. Importantly, this coordinated regulation of ROS by TaWRKY19 and TaWRKY40 enables wheat to have differential resistance against Pst races with diverse virulence levels.

In ascomycetes, perithecium development involves sexual differentiation processes regulated by mating-related signaling pathways and mating-type locus (MAT) transcription factors, activated by uncharacterized receptors in response to stage-specific signaling cues. Here, we show that a non-pheromone receptor, Gip1, regulates two distinct sexual differentiation processes during perithecial development in the wheat scab fungus Fusarium graminearum. Gip1 controls the formation of perithecium initials via the cAMP-PKA pathway, and regulates subsequent development, including the differentiation of peridia and ascogenous tissues, via the Gpmk1 MAPK pathway. The C-terminal tail of Gip1 is important for intracellular signaling, while its N-terminal region and extracellular loop 3 are key for ligand recognition. Interestingly, all sexual-specific spontaneous suppressors of gip1 had mutations in the FgVeA gene, encoding a component of the Velvet complex, which regulates sexual reproduction in filamentous ascomycetes. These mutations partially rescue defects in either perithecium initiation or maturation in gip1 mutants, and restore upregulation of genes important for perithecium development such as MAT1-1-2 (encoding a MAT transcription factor). Thus, Gip1 controls two early stages of sexual differentiation by activating downstream cAMP signaling and Gpmk1 pathways, which may coordinately regulate the expression of genes important for initial perithecium development via FgVeA.

Fusarium head blight (FHB), primarily caused by Fusarium graminearum, is a globally destructive fungal disease that not only reduces cereal crop yields but also threatens food safety due to mycotoxin contamination. In this study, we systematically characterized histone acetyltransferases in F. graminearum and revealed the overlapping functions between FgSas3 and FgRtt109. The double deletion mutant Fgsas3 Fgrtt109 showed severely impaired vegetative growth, conidiation, mycotoxin production, as well as complete loss of sexual reproduction and pathogenicity. Furthermore, integrated transcriptome and metabolome analyses revealed that this double mutant had significant dysregulation in carbohydrate metabolism, particularly in the disaccharides to monosaccharides conversion. This metabolic shift was evidenced by the reduced disaccharide concentrations, accumulated monosaccharide and their derivatives, and enhanced growth on disaccharide-supplemented medium in the Fgsas3 Fgrtt109 double mutant. Taken together, our results demonstrate that FgSas3 and FgRtt109 synergistically regulate carbohydrate metabolism, which in turn modulates fungal development, and plant infection.

The Rpd3 histone deacetylase complex is a multiple-subunit complex that mediates the regulation of chromatin accessibility and gene expression. Sin3, the largest subunit of Rpd3 complex, is conserved in a broad range of eukaryotes. Despite being a molecular scaffold for complex assembly, the functional sites and mechanism of action of Sin3 remain unexplored. In this study, we functionally characterized a glutamate residue (E810) in FgSin3, the ortholog of yeast Sin3 in Fusarium graminearum (known as wheat scab fungus). Our findings indicate that E810 was important for the functions of FgSin3 in regulating vegetative growth, sexual reproduction, wheat infection, and DON biosynthesis. Furthermore, the E810K missense mutation restored the reduced H4 acetylation caused by the deletion of FNG1, the ortholog of the human inhibitor of growth (ING1) gene in F. graminearum. Correspondingly, the defects of the fng1 mutant were also partially rescued by the E810K mutation in FgSin3. Sequence alignment and evolutionary analysis revealed that E810 residue is well-conserved in fungi, animals, and plants. Based on Alphafold2 structure modeling, E810 localized on the FgRpd3-FgSin3 interface for the formation of a hydrogen bond with FgRpd3. Mutation of E810 disrupts the hydrogen bond and likely affects the FgRpd3-FgSin3 interaction. Taken together, E810 of FgSin3 is functionally associated with Fng1 in the regulation of H4 acetylation and related biological processes, probably by affecting the assembly of the Rpd3 complex.

Eukaryotes have evolved defense systems to protect their genomes from invasive genetic elements. We report a fungal defense mechanism, tandem repeat-induced sexual silencing (TRISS), active during sexual stages. TRISS is a unique RNA interference (RNAi) pathway distinct from known mechanisms like quelling and meiotic silencing by unpaired DNA (MSUD). Triggered by tandem repeats, it involves RNAi components similar to quelling but uniquely includes the MSUD-specific Argonaute Sms2. TRISS operates independently of recombination. We identified small interfering RNA sassociated with TRISS (trasiRNAs) in tandem repeats. Sms2, guided by trasiRNAs, mediates translational repression of target mRNAs and is crucial for trasiRNA biogenesis. TRISS requires the DNA methyltransferase Rid but not Rid-mediated repeat-induced point mutation (RIP). Both Sms2 and Rid interact with replication protein A (RPA) to recruit the RPA-Qde1/RdRP complex, dependent on the helicase Qde3. This study reveals a potentially conserved strategy linking RIP and RNAi to silence tandem repeats during fungal sexual stages, offering insights into fungal genome defense.

Fusarium head blight (FHB) is a devastating wheat disease. Fhb1, the most widely applied genetic locus for FHB resistance, is conferred by TaHRC of an unknown mode of action. Here, we show that TaHRC alleles distinctly drive liquid-liquid phase separation (LLPS) within a proteinaceous complex, determining FHB susceptibility or resistance. TaHRC-S (susceptible) exhibits stronger LLPS ability than TaHRC-R (resistant), and this distinction is further intensified by fungal mycotoxin deoxynivalenol, leading to opposing FHB symptoms. TaHRC recruits a protein class with intrinsic LLPS potentials, referred to as an “HRC-containing hub.” TaHRC-S drives condensation of hub components, while TaHRC-R comparatively suppresses hub condensate formation. The function of TaSR45a splicing factor, a hub member, depends on TaHRC-driven condensate state, which in turn differentially directs alternative splicing, switching between susceptibility and resistance to wheat FHB. These findings reveal a mechanism for FHB spread within a spike and shed light on the roles of complex condensates in controlling plant disease.

ING proteins display a high level of evolutionary conservation across various species, and play a crucial role in modulating histone acetylation levels, thus regulating various important biological processes in yeast and humans. Filamentous fungi possess distinct biological characteristics that differentiate them from yeasts and humans, and the specific roles of ING proteins in filamentous fungi remain largely unexplored. In this study, an ING protein, Fng2, orthologous to the yeast Pho23, has been identified in the wheat head blight fungus Fusarium graminearum. The deletion of the FNG2 gene resulted in defects in vegetative growth, conidiation, sexual reproduction, plant infection, and deoxynivalenol (DON) biosynthesis. Acting as a global regulator, Fng2 exerts negative control over histone H4 acetylation and governs the expression of over 4000 genes. Moreover, almost half of the differentially expressed genes in the fng3 mutant were found to be co-regulated by Fng2, emphasizing the functional association between these two ING proteins. Notably, the fng2 fng3 double mutant exhibits significantly increased H4 acetylation and severe defects in both fungal development and pathogenesis. Furthermore, Fng2 localizes within the nucleus and associates with the FgRpd3 histone deacetylase (HDAC) to modulate gene expression. Overall, Fng2's interaction with FgRpd3, along with its functional association with Fng3, underscores its crucial involvement in governing gene expression, thereby significantly influencing fungal growth, asexual and sexual development, pathogenicity, and secondary metabolism.