RNA 5-methylcytosine (m5C) plays an undefined role in plant antiviral defense. Here, we reveal that the wheat methyltransferase NOP2/Sun RNA methyltransferase 2 (TaNSUN2) is an RNA m5C methyltransferase recruited by eukaryotic elongation factor 1-alpha (TaeEF1A) into Chinese wheat mosaic virus (CWMV) replication complexes (VRCs). TaNSUN2 promotes the m5C modification of CWMV RNAs to stabilize them and enhance their translational efficiency. Moreover, m5C modification in RNA 3 ' untranslated region strengthens both TaeEF1A binding, promoting viral replication, and viral coat protein interaction, facilitating viral assembly. A major allele of TaNSUN2 prevents its interaction with TaeEF1A and hinders its entry into the VRCs to suppress m5C modification of viral RNAs. TaNSUN2 knockout enhances wheat resistance to CWMV and increases the weight and size of wheat grains. Our findings provide mechanistic insights into the function of RNA modifications in plant virus infection and a valuable genetic resource for future wheat breeding projects.

Ferroptosis, an iron-dependent form of regulated cell death driven by lipid peroxidation, was initially characterized as a cell-autonomous process. However, emerging evidence demonstrates that ferroptotic signals can propagate between cells, triggering synchronized death events with significant pathophysiological implications. This review examines the molecular mechanisms underlying intercellular ferroptosis propagation, including ROS-mediated trigger waves, contact-dependent lipid peroxidation transmission, paracrine signaling, and extracellular vesicle-mediated communication. We analyze the dual roles of ferroptosis propagation in the tumor microenvironment and its contribution to tissue damage in ischemia-reperfusion injury and neurodegeneration. Understanding these propagation mechanisms offers novel therapeutic opportunities for precisely modulating cellular death responses across diverse disease contexts.

Alternaria brassicicola is a necrotrophic fungal pathogen causing black spot disease on cruciferous crops worldwide. In this study, we comprehensively characterized the pathogenic isolate Ab0920a from diseased broccoli in Shanghai using morphological, phylogenetic, host range, fungicide sensitivity, genomic, and functional analyses. Pathogenicity tests on 27 cruciferous varieties revealed a broad host range with varying resistance levels. Fungicide sensitivity assays showed that fluxapyroxad (EC50 = 0.0695 µg/mL), prochloraz (0.0711 µg/mL), and difenoconazole (0.0863 µg/mL) were highly effective, whereas fluazinam was least effective. Genome-wide annotation identified 941 secreted proteins (8.95% of the proteome) and 237 candidate effectors, including 31 small cysteine-rich secreted proteins and 68 homologs of known virulence factors. Conserved effector-associated motifs (e.g., RXLR, [Y/F/W]xC) were detected, and carbohydrate-active enzyme CAZyme annotation revealed diverse families potentially involved in plant cell wall degradation. Functional validation using the pSUC2 yeast system confirmed that N-terminal signal peptides of tested effectors are competent for secretion. A PVX-based transient expression assay in Nicotiana benthamiana identified two effectors that suppress Bax-induced programmed cell death, suggesting their potential roles in modulating host immunity. Overall, this study provides comprehensive insights into the pathogen Ab0920a, offering resources for disease management and functional studies on necrotrophic fungal pathogenesis.

Soil-borne wheat yellow mosaic virus (WYMV), transmitted by the Polymyxa graminis, poses a severe threat to global wheat production. Although seed microbiome is the first colonizer of the rhizosphere and root tissues, its role in defense against soil-borne WYMV remains unexplored. Here, using 12 wheat varieties with contrasting resistance to WYMV, we integrated seed microbiome characterization, culturable seed endophyte inoculation assays, hydroponic validation experiments and defense gene expression profiling, to elucidate the contribution of the seed microbiome to soil-borne viral resistance. We found that resistant varieties exhibited significantly reduced WYMV loads in both leaves (33.5

As a transcriptional co-factor of E2F, DP proteins are typically involved in the regulation of plant cell-cycle-related processes. However, whether DP proteins participate in transcriptional regulation associated with plant antiviral defence remains largely unclear. This study demonstrates that NbDPB, a DP family protein in Nicotiana benthamiana, positively regulates resistance to potato virus X (PVX) by functioning as a transcription factor. Knockout of NbDPB increased plant susceptibility to PVX, while its overexpression significantly suppressed viral accumulation. Furthermore, we identified that NbDPB binds to the promoter of NbMYB (a MYB transcription factor) and positively regulates its expression. Silencing NbMYB enhanced PVX infection, whereas exogenous application of jasmonic acid (JA) and salicylic acid (SA) partially rescued this phenotype. Hormone levels of JA and SA, as well as the expression levels of their marker genes, were significantly reduced in the silenced NbMYB plants. These findings suggest that the NbDPB-NbMYB module mediates antiviral defence through the JA and SA signalling pathways. This study reveals a previously uncharacterized role of DP proteins in antiviral transcriptional regulation, indicating that they may play a crucial role in plant antiviral defence.

Abstract Viral suppressors of RNA silencing (VSRs) are crucial for viral infection. Here, we show that a wheat ( Triticum aestivum ) oligosaccharyltransferase ( TaOST1B ) is associated with resistance to Wheat yellow mosaic virus (WYMV). TaOST1B interacts with WYMV-encoded P1 and enhances the VSR function of P1 by N-glycosylating its asparagine residue at position 116. This increases intranuclear accumulation of P1 through interaction with the nuclear transport protein TaIMP-α2 and blocks the interaction between calmodulin (CaM3) and CaM-binding transcriptional activator (CaMTA3) to suppress RNA interference. Nevertheless, nonglycosylated P1 loses its VSR function and forms aggregates triggering endoplasmic reticulum (ER) stress and is subsequently degraded by the 26S proteasome. A natural variant of TaOST1B fails to bind P1 and regulate its N-glycosylation, which induces ER stress and proteasome-mediated degradation to attenuate WYMV infection. Our study identifies an oligosaccharyltransferases as being utilized by VSR to promote viral infection, offering insights into the arms race between plants and viruses.

Long noncoding RNAs (lncRNAs) are increasingly recognized as sources of functional peptides that regulate plant development and immunity, yet little is known about whether lncRNA-encoded peptides participate in antiviral defense. Through transcriptomic analysis combined with peptide-coding potential prediction, we identified wheat lncRNA19864, which is highly expressed in wheat yellow mosaic virus (WYMV)-resistant cultivars and encodes a 39-amino-acid peptide, TaLEP1. Genetic manipulation of TaLEP1 expression revealed that TaLEP1 acts as a positive regulator of wheat resistance to WYMV. Mechanistically, TaLEP1 forms oligomers and interacts with both the host autophagy factor TaATG18a and the viral replicase NIb, thereby facilitating NIb recruitment to autophagosomes for degradation. Disruption of TaLEP1 oligomerization or TaATG18a function impaired NIb degradation and compromised TaLEP1-mediated antiviral resistance. Notably, TaLEP1 also confers resistance to other Potyviridae members, including turnip mosaic virus and soybean mosaic virus, by similarly targeting their NIb proteins for degradation. These findings demonstrate that an lncRNA-encoded peptide is involved in plant antiviral immunity and offer a potential strategy for engineering broad-spectrum virus resistance in crops.

Plant viruses pose a serious threat to the production of numerous economically valuable food and fuel crops worldwide. Exploring protein interaction networks between viruses and their hosts offers vital insights into the viral infection process and facilitates the development of innovative antiviral strategies for plants. In this review, we aimed to explore commonly used identification methods for protein‒protein interactions (PPIs) and the latest relevant advances, focusing on the advantages and limitations of various PPI identification methods, as well as the role of protein interaction networks involved in virus‒host interactions. This overview of plant‒virus interactions can significantly contribute to insights into viral pathogenesis and innovative strategies for plant antiviral defense.

Although RNA cytosine-5 methylation (m5C) is an important post-transcriptional regulatory mechanism, its contribution to plant antiviral immunity remains unclear. In this study, we identified Thiamine thiazole synthase 2 (TaTHI2) as a host mRNA target of the wheat m5C methyltransferase TaNSUN2 during infection by Chinese wheat mosaic virus (CWMV), a soil-borne virus that poses a major threat to wheat production. TaNSUN2 contributes to the m5C modification of TaTHI2 transcripts, enhancing mRNA stability and sustaining TaTHI2 accumulation. The disruption of a key m5C site markedly reduced methylation, weakened TaNSUN2–RNA binding, and accelerated transcript decay, leading to the compromised production of reactive oxygen species (ROS) and increased viral infection. Mechanistically, the TaNSUN2-dependent m5C modification stabilized TaTHI2 mRNA, thereby promoting ROS-mediated antiviral defense. Collectively, our results establish the m5C modification of TaTHI2 mRNA as a critical post-transcriptional control point in CWMV resistance and highlight TaNSUN2-dependent RNA methylation as an integral component of host antiviral immunity.

Long noncoding RNAs (lncRNAs) are increasingly recognized as a source of functional peptides involved in plant development and immunity, yet little is known about whether lncRNA-encoded peptides participate in antiviral defence. By transcriptome analysis with peptide-coding potential prediction, we identify lncRNA19864, which is highly expressed in wheat yellow mosaic virus (WYMV)-resistant cultivars and encodes a 39-amino-acid peptide, TaLEP1. Genetic manipulation of TaLEP1 expression showed that TaLEP1 as a positive regulator of wheat resistance to WYMV. Mechanistically, TaLEP1 forms oligomers and interacts with both the host autophagy factor TaATG18a and the viral replicase NIb, thereby facilitating NIb recruitment to autophagosomes for degradation. Disruption of TaLEP1 oligomerization or TaATG18a function impaired NIb degradation and compromised TaLEP1-mediated antiviral resistance. Notably, TaLEP1 also confers resistance to other Potyviridae members, including Turnip mosaic virus (TuMV) and Soybean mosaic virus (SMV), by similarly targeting their NIb proteins for degradation. Accordingly, lncRNA-encoded peptide is involved in plant antiviral immunity. Our results offer a potential strategy for engineering broad-spectrum virus resistance in crops.

The soil microbiome plays a crucial role in determining soil health and supporting sustainable crop production. However, the microbial characteristics, especially key functional microorganisms, between healthy and diseased soil are not fully understood yet. This study aims to elucidate the mechanisms by which beneficial microbes suppress disease in continuous cropping systems, using Chinese yam (Dioscorea polystachya Turcz.) as the target crop. To achieve this, rhizosphere microbial communities from healthy and diseased plants were first compared in a field trial. We found a significant (P < 0.05) reduction in fungal diversity in the rhizosphere soils of diseased plants and an increase in bacterial diversity within yam tuber tissues. Significant differences in the relative abundance of key taxa, i.e., Fusarium and Bacillus, were further observed between diseased and healthy samples. We then successfully isolated six culturable Bacillus strains from healthy samples with several beneficial traits. This included antagonism against the yam pathogen (Fusarium oxysporum SYJJ6), biofilm formation, and IAA production capacity. Subsequently, both pot and field experiments demonstrated that when inoculated into soil, isolated Bacillus strains (especially strain C21) significantly reduced the abundance of Fusarium spp. in the yam rhizosphere soil and increased biomass and yield. In conclusion, our study shows that healthy yam rhizosphere soils could suppress pathogen invasion and improve crop production via stimulating indigenous beneficial microorganism proliferations. Among them, indigenous Bacillus populations are among the key taxa responsible for maintaining yam rhizosphere health, and their re-inoculation effectively restores the disease-suppressive function of degraded soils. These findings highlight the importance of manipulating soil microbiome to enhance soil health, facilitating targeted crop management and boosting agricultural productivity.

Low temperature as one of the most important abiotic stresses has caused serious wheat production loss worldwide. We identified TaWRKY115 affecting cold tolerance in wheat by integration of genome-wide association study and RNA sequencing. Overexpression and CRISPR/Cas9-mediated gene-editing revealed TaWRKY115 positively modulating cold tolerance in wheat. Transcription factor TaSP1 negatively regulates the cold tolerance of wheat by binding to the promoter of TaWRKY115 in cold-sensitive haplotypes. TaWRKY115 inhibited TaMYB4 at both protein and transcription levels. BSMV-mediated silenced wheat plants and EMS mutants of TaMYB4 possessed increased wheat cold tolerance. We further revealed that TaMYB4 repressed expressions of the TaCBF family through binding to their promoters. BSMV-mediated silenced wheat plants and EMS mutants of TaCBF12d exhibited decreased cold tolerance in wheat. This study brought forth a new model TaSP1-TaWRKY115-TaMYB4, regulating wheat cold tolerance through the TaCBF pathway and provided valuable cold tolerance genes for wheat breeding programs.

Early mutualistic interactions between host plants and their rhizosphere microbes have the potential to provide soil-borne disease resistance. However, it remains unclear how the early rhizosphere microbiome protects against viral diseases such as wheat yellow mosaic virus, which is a major threat to global wheat production. We combined field trials with microbiome transplantation experiments to investigate the role of early rhizosphere microbiomes in suppressing wheat yellow mosaic disease. To uncover the underlying mechanisms, we further performed integrated multi-omics analyses of microbial communities, functional genes, and metabolic profiles. Disease-resistant wheat cultivars were consistently associated with distinct seedling rhizosphere microbiome assembly, including a lower Polymyxa graminis abundance, lower community compositional variation, and enrichment of beneficial taxa such as Bacillus, Pseudomonas, and Trichoderma. Resistant cultivars also exhibited distinct rhizosphere metabolite profiles, including higher levels of glyceraldehyde and N-acetyltryptophan, which were positively associated with keystone microbial taxa and stimulated representative isolates in vitro. Isolate-based and synthetic community validation further supported the functional relevance of these taxa, while microbial inoculation was associated with reduced vector abundance, lower virus accumulation, and activation of host defense-related pathways. Our findings showed that early cultivar-dependent rhizosphere microbiome assembly was closely linked to resistance against soil-borne viral disease in wheat.

Reactive oxygen species (ROS), as signaling molecules, play a crucial role in the plant immune response. However, the mechanism(s) by which viruses affect ROS metabolism remain largely unexplored. Here, we found that wheat yellow mosaic virus (WYMV)-encoded P1 is a pathogenic protein. Transcriptomic and proteomic integrative analyses were performed on WYMV-infected overexpressing-P1 wheat and wild-type plants. A total of 9245 differentially expressed genes (DEGs) and 1383 differentially expressed proteins (DEPs) were identified in the transcriptome and proteome, respectively. At their intersection, 373 DEGs/Ps were identified. Enrichment analysis revealed that the expression of genes related to the ROS metabolism pathway in overexpressed P1 transgenic wheat (OE-P1) plants significantly increased during WYMV infection. We screened peroxidase (TaPOD) and thioredoxin reductase (TaTrxR) as they showed the most significant differences in expression. The silencing of TaPOD and TaTrxR revealed that they positively regulate WYMV infection by reducing ROS accumulation. Furthermore, hydrogen peroxide treatment induced WYMV resistance in wild-type wheat plants and OE-P1 transgenic plants. This study provides a theoretical basis for the role of P1 in plant viral infection.

Ym1 is the most widely utilized gene for wheat yellow mosaic virus (WYMV) disease control in worldwide wheat breeding. Here, we successfully isolated the responsible gene for Ym1. It encodes a typical CC-NBS-LRR type R protein, which is specifically expressed in root and induced upon WYMV infection. Ym1-mediated WYMV resistance is likely achieved by blocking viral transmission from the root cortex into steles, thereby preventing systemic movement to aerial tissues. Ym1 CC domain is essential for triggering cell death. Ym1 specifically interacts with WYMV coat protein, and this interaction leads to nucleocytoplasmic redistribution, a process for transitioning Ym1 from an auto-inhibited to an activated state. The activation subsequently elicits hypersensitive responses and establishes WYMV resistance. Ym1 is likely introgressed from the sub-genome Xn or Xc of polyploid Aegilops species. The findings highlight an exogenous-introgressed and root-specifically expressed R gene that confers WYMV resistance by recognizing the viral component.

Papain-like cysteine proteases (PLCPs) play critical roles in regulating plant immunity against a range of pathogens and a series of cysteine protease inhibitors have been identified, however, relatively little research has been done on proteins that enhance the protease activity of PLCPs. Here, we identified a protein named NbPIRIN, the silencing of NbPIRIN promotes Chinese wheat mosaic virus (CWMV) infection, whereas the transgenic overexpression of NbPIRIN inhibits CWMV infection in Nicotiana benthamiana. Furthermore, we found that NbPIRIN interacts with papain-like cysteine protease (NbRD21) and increases its protease activity. We demonstrated that the silencing of NbRD21 significantly increased host susceptibility to CWMV infection, whereas the transgenic overexpression of NbRD21 increased host resistance. Interestingly, CWMV CRP was found to interact with both NbPIRIN and NbRD21, thus interfering with the interaction between NbPIRIN and NbRD21 and subsequently inhibiting the protease activity of NbRD21. Since wheat is the natural host of CWMV, we identified TaPIRIN and TaRD21 and found that they had functions similar to those of NbPIRIN and NbRD21 in the CWMV response. These results reveal a previously unreported offensive and defensive strategy between plants and viruses.

Posttranscriptional and posttranslational modifications play crucial roles in plant immunity. However, how plants fine-tune such modifications to activate antiviral immunity remains unknown. Here, we report that the m6A methyltransferase TaHAKAI is utilized by wheat yellow mosaic virus (WYMV) to increase viral genomic m6A modification and promote viral replication. However, TaHAKAI also functions as an E3 ligase that targets the viral RNA silencing suppressor P2 for degradation and inhibits viral infection. A major allele of TaHAKAI in a susceptible cultivar exhibited reduced E3 ligase activity but not m6A methyltransferase activity, promoting viral infection. Interestingly, TaHAKAIR attenuates the stability of TaWPS1 (Wheat paired spikelets 1, WPS1) mRNA, the negative regulator of spike development, which might increase panicle length and spikelet number by modulating its m6A modification. Our study reveals a mechanism for balancing disease resistance and yield by fine-tuning m6A modification and ubiquitination.

In plants, small peptides are important players in the plant stress response, yet their function in plant antiviral responses remains poorly understood. Here, we identify that the plant small peptide, CLAVATA3/ESR-RELATED 7 (CLE7), enhances plant resistance to Chinese wheat mosaic virus infection in Nicotiana (N.) benthamiana. Subsequent investigations demonstrate that CLE7 recognizes receptor kinase NbPXC3 to control the plant antiviral response. Moreover, CLE7-NbPXC3 signaling induces NbMKK2-controlled NbMPK4 phosphorylation, resulting in phosphorylation of the transcription factor NbEDT1. NbEDT1 phosphorylation is involved in the transcriptional activity of NbNCED3, which is a rate-limiting enzyme in abscisic acid (ABA) biosynthesis. Moreover, CLE7 activates broad-spectrum disease resistance to multiple RNA viral infections. Our study indicates that CLE7 induces a plant antiviral response through a series of immune signal transductions in N. benthamiana and provides a foundation for the exploration of efficient viral disease management methods based on plant small peptides.

Soil microbial alpha diversity is essential for driving ecosystem functions and processes. However, little is known about the beta-diversity affect community functions. Here, we combine distinct community inocula using the dilution-to-extinction approach with two wheat genotypes to study the effect of microbial diversity loss on rhizosphere community assembly processes, which are related to beta-diversity (between-habitat diversity), and the consequences for ecosystem functions within greenhouse experiment. Compared with alpha-diversity, the bacterial and fungal community beta-diversity are stronger predictors of ecosystem functions (organic matter degradation, phosphorus supply capacity and nitrogen supply capacity), plant genotypes regulated the relationship between microbial diversity and ecosystem functions, with ecosystem functions being significant link to microbial diversity under different wheat genotypes. Loss of microbial diversity decreased the abundance of Bacterial_ASV6 (Burkholderia) and increased Fungal_11 (Altemaria) within the restored rhizosphere soil. Null modeling analysis showed that the deterministic assembly processes are dominant in bacterial community and fungal high-diversity (alpha-diversity) community, associating with the change of specialized functions (organic matter degradation, phosphorus supply capacity and nitrogen supply capacity) that are correlated with microbial diversity and specific microbial taxa. In addition, these two species were key role for regulating to the network cohesion. Overall, our study pointed out that the regulation of community assembly by microbial diversity loss limits the development of soil ecological functions and weakens the stability of rhizosphere microbial network, highlighting the potential regulatory effect of microbial taxa distribution on microbial community stability and changes of specific ecological functions.