This study presents the synthesis of a novel nanocomposite, denoted as Pt/Se-TiO2@CNTs, which comprises platinum (Pt)-decorated selenium-doped titanium dioxide (Se-TiO2) supported on functionalized carbon nano-tubes (FCNTs). The synthesis was performed using a sol-gel method, facilitating the in-situ decoration of platinum nanoparticles (NPs) for uniform dispersion and robust interfacial connections. Characterization techniques, including X-Ray Diffraction (XRD), Transmission Electron Microscopy (TEM), Scanning Electron Microscopy equipped with Energy Dispersive Spectroscopy (SEM-EDS), Fourier Transform Infrared Spectroscopy (FTIR), Raman Spectroscopy, UV-visible spectroscopy, and X-Ray Photoelectron Spectroscopy (XPS), confirmed the incorporation of Se into the TiO2 and the successful decoration of Pt, along with enhanced optical absorption in the visible spectrum. The nanocomposite functioned as an effective photocatalyst, achieving a 95.4% degradation of indigo carmine dye within 120 min. In vitro experiments showed that the nanocomposite at a concentration of 300 & micro;g/L demonstrated 85% colony growth inhibition of Ralstonia solanacearum at various time intervals (20, 40, 60, 80, 100, 120, and 140 h) under visible light, indicating significant photocatalytic antibacterial activity. The nanocomposite was then applied to tomato plants via foliar spray at concentrations of 100, 200, and 300 mg/L, with the 300 mg/L concentration proving most effective in mitigating bacterial wilt disease, thus enhancing physiological and morphological parameters including fresh weight (19.70 g), dry weight (3.36 g), shoot length (30.40 cm), root surface area (25.26 cm), and plant length (33.90 cm). Furthermore, it improved total chlorophyll (2.10 mg/g) and carotenoid contents (1.85 mg/g) at this concentration. The biochemical application also significantly elevated antioxidant enzyme activities like superoxide dismutase (SOD) (74 mg/g), peroxidase (POD) (22.6 mg/g), catalase (CAT) (26.9 mg/g), and ascorbate peroxidase (APX) (21.7 mg/g), along with increased phenolic and flavonoid contents. This research highlights the potential of Pt-decorated Se-TiO2@CNTs nanocomposites for effective antibacterial applications against plant pathogens and their environmental degradations.

Plant extracellular vesicles (EVs) serve as critical mediators of intercellular communication during plant-pathogen interactions, particularly through their cargo of regulatory small RNAs, enabling the transport of miRNAs to distant tissues during biotic stress. Potato virus Y (PVY), one of the most economically damaging plant viruses globally, poses significant threats to solanaceous crop production. However, the landscape of EV-associated miRNAs and their regulatory roles in PVY infection remain largely unexplored. In this study, we isolated and characterized EV-associated particles from the apoplastic fluid of both PVY-infected and healthy tomato leaves using differential ultracentrifugation, followed by transmission electron microscopy, nanoparticle size analysis, and western blotting. High-throughput small RNA sequencing revealed 96 significantly differentially expressed miRNAs in EV-associated particles upon viral challenge. Bioinformatic prediction revealed that 80% of these dysregulated miRNAs potentially target multiple genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses demonstrated significant overrepresentation of predicted target genes in pathways associated with transcription, ta-siRNA biogenesis involved in RNA interference, protein binding, RNAi-mediated antiviral immune response, oxidative phosphorylation, mRNA surveillance pathway, and eukaryotic ribosome biogenesis. Our findings demonstrate that PVY infection selectively modulates the miRNA composition within tomato EV-associated particles. These EV-associated particles delivered miRNAs may contribute to a sophisticated antiviral defense mechanism by co-regulating host immunity. This study provides novel insights into the role of EV-associated particles mediated RNA communication in plant immunity and lays a theoretical foundation for developing innovative miRNA- and EV-based antiviral strategies for crop protection.

INTRODUCTION:Synthetic nitrogen (N) fertilizers underpin global food security but drive escalating environmental costs, including greenhouse gas (GHG) emissions and soil degradation. Conventional N-reduction strategies face a fundamental trade-off: lowering fertilizer inputs typically compromises crop productivity and farmer income. Nanotechnology offers a pathway to decouple yield from N dependency, yet scalable, low-cost solutions remain elusive. OBJECTIVES:This study aimed to develop a least-input foliar fertilization strategy using bio-derived nanocarbon (BNC) that enables a 30% reduction in synthetic N input in peanut (Arachis hypogaea L.) without sacrificing crop yield, nutritional quality, or economic returns, while elucidating the underlying physiological and microbiological mechanisms. METHODS:Field trials were conducted with peanut (cv. Huayu 22) over two consecutive growing seasons (2024-2025) in Laixi, Shandong, China, under four treatments: N (conventional N, 100% urea), N-30 (reduced N, 70% urea), N + BNC (100% urea + foliar BNC), and N-30 + BNC (70% urea + 18 g ha-1 foliar BNC). Photosynthetic parameters, oxidative stress markers, and 15N isotope tracing were assessed. Transcriptomic, metabolomic, and 16S rRNA microbiome analyses characterized systemic signaling and rhizosphere responses. GHG emissions were modeled using the DNDC framework, and economic benefits were calculated based on yield and input costs. RESULTS:BNC application under N-30% maintained yield parity with the N control by sustaining photosynthetic electron transport and reducing reactive oxygen species (ROS). This enhanced carbon status upregulated the transcription factor HY5, activating nitrate transporter AhNRT1.2 and ammonium transporter AhAMT1.1 and increasing 15N uptake. Changes in root-exudate composition were accompanied by shifts in the rhizosphere bacterial community, including higher relative abundance of taxa associated with nitrification and nutrient turnover (Nitrospira and Gemmatimonas), and coincided with 21% and 14% higher root NO3--N and NH4+-N contents, respectively. Nitrogen use efficiency (NUE) improved by 16.0%, total GHG emissions decreased by 34.8%, and net profit increased by 27.0%. CONCLUSION:A micro-dosage of upcycled BNC (∼$0.02 ha-1) orchestrates HY5-mediated shoot-to-root signaling and rhizosphere microbiome restructuring to sustain crop productivity under reduced N inputs. This commercially viable, low-input framework offers a scalable route for climate-smart agriculture that reconciles food security with environmental sustainability.

Cytosine-5 methylation (m5C) is a crucial epitranscriptomic mark in eukaryotes that modulates RNA stability and gene expression. While the roles of m5C are partially understood in model plants, its function in horticultural crops under biotic stress remains largely unexplored. To address this gap, we investigated the role of m5C modification in tomato response to tomato spotted wilt virus (TSWV) infection. We constructed the first comprehensive m5C epitranscriptomic map in Solanum lycopersicum. To investigate its role in plant immunity, we further profiled the dynamic changes of the m5C methylome upon TSWV infection, followed by integrative multi-omics analysis. Functional validation was performed through virus-induced gene silencing (VIGS) of the key RNA methyltransferase gene SlTRM4B. The m5C epitranscriptomic map revealed conserved modification patterns with enrichment at transcription start and stop sites. Upon TSWV infection, a global increase in m5C modification levels was observed across the transcriptome, which was correlated with the significant upregulation of RNA methyltransferase (RCMT) family genes, particularly SlTRM4B. Integrative multi-omics analysis revealed that genes exhibiting both hypermethylation and increased expression were significantly enriched in the plant-pathogen interaction pathway. VIGS of SlTRM4B demonstrated that this methyltransferase is essential for maintaining the stability of its target transcripts under TSWV infection, leading to enhanced disease susceptibility. Collectively, our findings demonstrate that SlTRM4B-mediated m5C RNA methylation fine-tunes post-transcriptional regulation to reprogram the transcriptome for disease resistance in tomato. This work provides novel insights into the epitranscriptomic mechanisms governing plant responses to viral pathogens.

A systematic elucidation of pathogenic factors and molecular evolution in tobacco leaf pathogens is pivotal for green precision prevention and control. In this study, isolate A2 was obtained from diseased tobacco leaves, and its strong pathogenicity to tobacco was confirmed through pin-prick inoculation following Koch’s postulates. Phylogenetic analysis, average nucleotide identity (ANI), and digital DNA-DNA hybridization (dDDH) results clearly identified A2 as Pseudomonas monteilii. Whole-genome sequencing revealed that A2 is enriched in genes associated with metabolic regulation, environmental adaptation, and virulence. Gene families linked to pathogenic potential, including acetyltransferases, phosphatidylserine-associated proteins, two-component heavy metal response transcriptional regulators, von Willebrand factor type A proteins, TIGRFAM RND efflux system outer membrane lipoprotein NodT, and multicopper oxidoreductases, exhibited significant expansion or contraction, suggesting optimization of host adaptability. Integrated multidimensional annotations from the Virulence Factor Database (VFDB), Pathogen-Host Interactions Database (PHI-base), and Carbohydrate-Active Enzymes Database (CAZy) indicate that A2 pathogenicity is driven by a composite “structure–metabolism–regulation” network involving motility-related structures, secretion systems, and polysaccharide metabolism, with the lipopolysaccharide biosynthesis gene waaF serving as a critical node for outer membrane stability and immune evasion. In summary, this is the first confirmed P. monteilii isolate pathogenic to tobacco, expanding the known host range of this species and providing novel genomic insights into its phytopathogenic mechanisms.

Lysinibacillus fusiformis strain KBD-5, previously known for its antiviral activity against Tobacco mosaic virus, was investigated for its biocontrol potential against the fungal pathogen Botrytis cinerea. In plate assays, conducted with three independent biological replicates and incubated at 28 °C for 5 days, KBD-5 significantly inhibited the mycelial growth of B. cinerea by 76.42

Melatonin, a pleiotropic regulatory factor, plays a key role in mediating crop drought resistance. Herein, we conducted an integrated physiological and transcriptomic approach to elucidate the mitigating effect of endogenous melatonin in mitigating drought stress in maize. We generated a comt snat asmt maize mutant via CRISPR-Cas9-mediated simultaneous editing of ZmCOMT, ZmSNAT, and ZmASMT1-genes encoding rate-limiting enzymes in the endogenous melatonin biosynthesis pathway. Sequencing of the mutant lines revealed key amino acid substitutions (Gly168→Ala in ZmSNAT, Asp175→Glu in ZmCOMT, and Asp150→Glu in ZmASMT1) within critical protein domains, resulting from CRISPR-induced small insertions or deletions (indels), which led to subtle alterations in the tertiary conformation of corresponding proteins. These modifications resulted in an 86.70% increase in endogenous melatonin content. Under drought stress, the comt snat asmt maize exhibited enhanced antioxidant enzyme activities, leading to a significant reduction in reactive oxygen species (ROS) accumulation compared to the control. Furthermore, endogenous levels of melatonin, abscisic acid (ABA), cytokinin (CTK), and auxin (IAA) were markedly elevated, whereas gibberellin (GA) content was significantly reduced. Consistently, the activities of SNAT, ASMT, and COMT were also enhanced in the mutant. Transcriptomic profiling further revealed that endogenous melatonin regulates ABA, IAA, CTK, and GA signaling pathways to enhance drought tolerance. In particular, ZmCOMT, ZmSNAT, and ZmASMT1 apparently modulated the expression levels of key regulatory genes such as ZmIAA2, ZmIAA23, ZmIAA7, ZmSAUR24, ZmPYL8, and ZmPIF3.1, associated with these hormone pathways. Collectively, endogenous melatonin reinforces drought tolerance by reducing ROS accumulation and reprogramming phytohormone homeostasis through regulation of hormone-related gene expression. Our findings provide important insights into the regulatory mechanisms by which endogenous melatonin enhances drought resistance in crops.

The valorization of agricultural waste into functional nanomaterials represents a critical pathway toward sustainable agriculture. In this study, we employed flash Joule heating (FJH) technology to upcycle discarded tea tree straw into flash graphene (FG). This solvent-free, ultra-fast process yielded a turbostratic carbon material with layered morphology. The synthesized FG exhibited a 2D layered structure and an average particle size of 14.15 ± 0.6 µm, as characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), and Raman spectroscopy. Foliar application of FG reduced PVY-associated symptoms and viral accumulation in Nicotiana benthamiana. Transcriptomic analysis revealed route-dependent changes in photosynthesis- and defense-associated programs. NbWRKY40 was significantly upregulated following FG exposure, and gain- and loss-of-function experiments supported its positive role in antiviral immunity. Bioinformatic analysis and live-cell imaging identified predicted disordered/prion-like features and dynamic nuclear condensate-like structures. These observations are consistent with a possible role for biomolecular condensation, but do not by themselves establish bona fide LLPS or direct induction by FG.

Background The practice of tobacco monoculture usually leads to increased occurrence of soil-borne diseases and reduced yield and quality, posing a significant obstacle to sustainable farming. The soil microbiome is central to soil health, but the mechanisms by which crop rotation alleviates monoculture-related obstacles by reconstructing microbial communities and their interaction networks remain poorly understood.Methods A field study was conducted in two regions to compare tobacco grown on soils after rotation (with wheat or rapeseed) and tobacco grown on soils with continuous tobacco cropping. A comprehensive analysis was performed on soil physicochemical properties, enzyme activities, microbial biomass, 16S rRNA bacterial and ITS fungal gene sequencing, microbial co-occurrence networks and phenotypic data, including tobacco yield, disease index and chemical quality of cured tobacco. A multi-omics coupling framework was constructed to integrate environmental variables, community structure and phenotype.Results Crop rotation notably increased the content of available potassium in the soil and enhanced the activities of key enzymes involved in carbon and phosphorus cycling, such as beta-glucosidase, sucrase and acid phosphatase. These changes led to improved nutrient availability. Across the two regions, crop rotation consistently restructured both bacterial and fungal communities, enriching beneficial taxa, including Chitinophagaceae, Rhodanobacteraceae, Xanthobacteraceae, Cercophora and Montagnula, while suppressing the proliferation of potential fungal pathogens. The microbial co-occurrence networks under rotation conditions were more intricate, cooperative and functionally integrated. Path analysis, incorporating composite variables, revealed a comprehensive causal pathway: crop rotation enhances the abundance of beneficial microbes and increases network complexity, which, in turn, improves the soil health indexes. This ultimately results in higher tobacco yield, reduced disease incidence and superior chemical and sensory quality of the cured tobacco.

Global warming poses a considerable threat to crop production, making heat stress a pivotal challenge in agriculture. Yet how epitranscriptomic modifications contribute to plant heat stress responses remains to be explored. Here, this study reveals the critical role of phase separation in plant heat stress tolerance and demonstrated that N-acetyltransferase 10 (NAT10), which encodes of the cytosine N4 acetyltransferase protein, contributes to heat resistance. We found that NAT10 interacts with polyadenylate-binding protein (PABP), which contains intrinsically disordered regions (IDRs), thereby facilitating the selective recruitment of ac4C-modified mRNAs into PABP-mediated condensates. Integrative transcriptome-wide analysis, combining ac4C acetylome profiling with SG-enriched transcript sequencing, revealed that detoxification-related mRNAs, including those encoding the cytochrome P450, phenylalanine ammonia-lyase, glutathione S-transferase, and heat shock 70 protein families, preferentially accumulate within these condensates. This accumulation maintains their stability and prevents stress-induced degradation. Conversely, loss of PABP impairs the recruitment of ac4C-modified detoxification-related transcripts into stress granules, thereby promoting their degradation under heat stress. In summary, our findings identify a stress-responsive NAT10-PABP-ac4C axis that promotes phase separation to stabilize ac4C-modified mRNAs under heat stress. By recruiting detoxification-related transcripts into stress granules, this axis ensures mRNA stability and offers insights for enhancing crop resilience under environmental stress.

N4-acetylcytidine (ac4C), which is an evolutionarily conserved RNA modification in eukaryotes, functions as a critical epitranscriptomic regulator that enhances mRNA stability and translation efficiency. In Arabidopsis thaliana and Oryza sativa, ac4C has been characterized as a modulator of transcriptome homeostasis that functions by regulating translational efficiency and maintaining RNA structure; however, the distribution patterns and biological functions of ac4C in other plant species remain largely unexplored. A transcriptome-wide map of the ac4C of Solanum lycopersicum revealed conserved features of plant ac4C modification, with preferential enrichment in mRNA translation initiation and termination regions as well as high conservation of modification motifs and functional categories of target genes across species. Notably, heat stress triggered global ac4C hyperacetylation, and hypermodified genes were enriched in transcripts encoding photosynthesis and thermotolerance-associated regulators. Multiomics integration further revealed that hyperacetylated and upregulated genes were significantly enriched in stress-response pathways, accompanied by elevated transcription levels. Silencing of SLNAT10 resulted in reduced stability of highly acetylated transcripts under heat stress conditions. These findings reveal that RNA acetylation is synchronized with posttranscriptional regulation that promotes transcriptional reprogramming to establish heat tolerance, providing a mechanism for the epitranscriptomic control of environmental adaptation in plants.

Infectious wound healing has garnered significant attention due to the increasing prevalence of drug-resistant bacteria, with photodynamic therapy (PDT) emerging as a promising non-invasive approach. Among PDT techniques, visible-light-activated therapies hold great potential. Nevertheless, current applications face critical challenges, such as non-specific targeting of bacteria, limited photodynamic efficacy, and insufficient oxygen supply. To resolve these limitations, the study introduces an innovative dual-functional bioinspired nanoparticle system. The aptamer-modified macrophage membranes enable high specificity and selectivity of the hybrid BiVO4/BiOI toward S. aureus. Simultaneously, the heterojunction formed between BiVO4 and BiOI quantum dots not only expands the photocatalytic active surface area but also significantly accelerates the separation of photogenerated charge carriers, thereby enhancing PDT activity. By utilizing visible-light irradiation to catalyze H2O molecules, the system generates reactive oxygen species (ROS) independent of ambient oxygen levels, overcoming the limitation of oxygen content availability in traditional PDT approaches. Computational simulations further confirm that the generated ROS effectively disrupt bacterial phospholipid bilayers, enhancing bactericidal efficacy. A series of biological evaluations, including in vitro and in vivo antibacterial assays and cell migration studies, validate the dual-functional of the bioinspired nanoparticles in accelerating wound healing. Overall, this multifunctional bioinspired nanocomposite, with its targeted delivery, visible-light responsiveness, and potent antibacterial properties, demonstrates substantial promise for applications in treating infectious wounds. STATEMENT OF SIGNIFICANCE: Infectious wound healing has garnered significant attention due to the increasing prevalence of drug-resistant bacteria, with photodynamic therapy emerging as a promising non-invasive approach. The heterostructure formed between BiVO4 and BiOI QDs exhibits highly efficient photocatalytic activity, which maximizes ROS production, leading to more effective bacterial eradication. To date, this specific heterostructure has not been extensively studied. Moreover, the aptamer-modified macrophage cell membranes achieve high specificity and selectivity of the hybrid BiVO4/BiOI toward S. aureus. This multifunctional bioinspired nanocomposite, with its targeted delivery, visible-light responsiveness, and potent antibacterial properties, demonstrates substantial promise in medical field.

The necrotrophic phytopathogen Rhizoctonia solani causes leaf spot and root rot in tobacco, which significantly reduce the yield and quality of the crop. Recently, much attention has been paid to R. solani effectors, but only a few of them have been characterized. Here, we identified a CAP domain-containing protein, RsCAP3, with a functional signal peptide, which triggers slight cell death on tobacco leaves. Meanwhile, RsCAP3 dramatically induces salicylic acid (SA) signaling but inhibits jasmonic acid (JA) signaling, increasing the plant's susceptibility to R. solani. Moreover, we demonstrated that RsCAP3 interacts with Nb14-3-3b, a negative regulator of tobacco resistance to R. solani. Correspondingly, we also found that transient expression of Nb14-3-3b activates SA signaling while suppressing JA signaling. Based on these results, we propose that RsCAP3 hijacks Nb14-3-3b to disrupt JA and SA pathways, thereby facilitating the infection of R. solani. Our findings highlight a novel virulence strategy in which R. solani interferes with the JA-/SA-mediated defense response.

Background Viral diseases continue to pose a major threat to the world’s commercial crops. The in-depth exploration and efficient utilization of resistance proteins have become crucial strategies for their control. However, current delivery methods for introducing foreign DNA suffer from host range limitations, low transformation efficiencies, tissue damage, or unavoidable DNA integration into the host genome. The nanocarriers provides a convenient channel for the DNA delivery and functional utilization of disease-resistant proteins. Results In this research, we identified a cysteine-rich venom protein (NbCRVP) in Nicotiana benthamiana for the first time. Virus-induced gene silencing and transient overexpression clarified that NbCRVP could inhibit the infection of tobacco mosaic virus, potato virus Y, and cucumber mosaic virus, making it a broad-spectrum antiviral protein. Yeast two-hybrid assay, co-immunoprecipitation, and bimolecular fluorescence complementation revealed that calcium-dependent lipid-binding (CaLB domain) family protein (NbCalB) interacted with NbCRVP to assist NbCRVP playing a stronger antiviral effect. Here, we demonstrated for the first time the efficient co-delivery of DNA expressing NbCRVP and NbCalB into plants using poly(amidoamine) (PAMAM) nanocarriers, achieving stronger broad-spectrum antiviral effects. Conclusions Our work presents a tool for species-independent transfer of two interacting protein DNA into plant cells in a specific ratio for enhanced antiviral effect without transgenic integration, which further demonstrated new strategies for nanocarrier-mediated DNA delivery of disease-resistant proteins. Graphical abstract

Potato virus Y (PVY) disease is a global problem that causes significant damage to crop quality and yield. As traditional chemical control methods are ineffective against PVY, it is crucial to explore new control strategies. MicroRNAs (miRNAs) play a crucial role in plant and animal defense responses to biotic and abiotic stresses. These endogenous miRNAs act as a link between antiviral gene pathways and host immunity. Several miRNAs target plant immune genes and are involved in the virus infection process. In this study, we conducted small RNA sequencing and transcriptome sequencing on healthy and PVY-infected N. benthamiana tissues (roots, stems, and leaves). Through bioinformatics analysis, we predicted potential targets of differentially expressed miRNAs using the N. benthamiana reference genome and the PVY genome. We then compared the identified differentially expressed mRNAs with the predicted target genes to uncover the complex relationships between miRNAs and their targets. This study successfully constructed a miRNA-mRNA network through the joint analysis of Small RNA sequencing and transcriptome sequencing, which unveiled potential miRNA targets and identified potential binding sites of miRNAs on the PVY genome. This miRNA-mRNA regulatory network suggests the involvement of miRNAs in the virus infection process.

The ubiquitin–proteasome system (UPS) plays an important role in virus–host interactions. However, the mechanism by which the UPS is involved in innate immunity remains unclear. In this study, we identified a novel major latex protein-like protein 43 (NbMLP43) that conferred resistance to Nicotiana benthamiana against potato virus Y (PVY) infection. PVY infection strongly induced NbMLP43 transcription but decreased NbMLP43 at the protein level. We verified that B-box zinc finger protein 24 (NbBBX24) interacted directly with NbMLP43 and that NbBBX24, a light responsive factor, acted as an essential intermediate component targeting NbMLP43 for its ubiquitination and degradation via the UPS. PVY, tobacco mosaic virus, (TMV) and cucumber mosaic virus (CMV) infections could promote NbMLP43 ubiquitination and proteasomal degradation to enhance viral infection. Ubiquitination occurred at lysine 38 (K38) within NbMLP43, and non-ubiquitinated NbMLP43(K38R) conferred stronger resistance to RNA viruses. Overall, our results indicate that the novel NbMLP43 protein is a target of the UPS in the competition between defense and viral anti-defense and enriches existing theoretical studies on the use of UPS by viruses to promote infection.

Nanoparticles (NPs) derived from RNA interference (RNAi) are considered a potentially revolutionary technique in the field of plant protection in the future. However, the application of NPs in RNAi is hindered by the conflict between the high cost of RNA production and the large quantity of materials required for field application. This study aimed to evaluate the antiviral efficacy of commercially available nanomaterials, such as chitosan quaternary ammonium salt (CQAS), amine functionalized silica nano powder (ASNP), and carbon quantum dots (CQD), that carried double-stranded RNA (dsRNA) via various delivery methods, including infiltration, spraying, and root soaking. ASNP-dsRNA NPs are recommended for root soaking, which is considered the most effective method of antiviral compound application. The most effective antiviral compound tested was CQAS-dsRNA NPs delivered by root soaking. Using fluorescence, FITC-CQAS-dsCP-Cy3, and CQD-dsCP-Cy3 NPs demonstrated the uptake and transport pathways of dsRNA NPs in plants when applied to plants in different modes. The duration of protection with NPs applied in various modes was then compared, providing references for evaluating the retention period of various types of NPs. All three types of NPs effectively silenced genes in plants and afforded at least 14 days of protection against viral infection. Particularly, CQD-dsRNA NPs could protect systemic leaves for 21 days following spraying.

IntroductionKac is a model for all acylation modification studies. Kac plays a critical role in eukaryotes and prokaryotes. It is mainly involved in six major biological functions: gene expression, signal transduction, cell development, protein conversion, metabolism, and metabolite transport.MethodWe investigated and compared the acetylation modification of proteins in healthy and tomato spot wilt virus (TSWV)-infected Nicotiana benthamiana leaves.ResultWe identified 3,418 acetylated lysine sites on 1962 proteins acetylation of proteins in the TSWV-infected and control groups were compared; it was observed that 408 sites on 294 proteins were upregulated and 284 sites on 219 proteins (involved in pentose phosphate, photosynthesis, and carbon fixation in photosynthesis) were downregulated after the infection. Overall, 35 conserved motifs were identified, of which xxxkxxxxx_K_ Rxxxxxxxxx represented 1,334 (31.63%) enrichment motifs and was the most common combination. Bioinformatic analysis revealed that most of the proteins with Kac sites were located in the chloroplast and cytoplasm. They were involved in biological processes, such as cellular and metabolic processes.DiscussionIn conclusion, our results revealed that Kac may participate in the regulation of TSWV infection in N. benthamiana.

Most crop viruses are carried and spread by seeds. Virus-infected seeds are seed-borne viral disease infections, and thus, reducing the rate of seed infection is an urgent problem in the seed-production industry. The objective of this study was to use nanoparticles (NPs) to directly deliver dsRNA into plants or pollen to initiate RNA interference (RNAi) to reduce viral carryover in seeds. Chitosan quaternary ammonium salt (HACC), complexed with dsRNAs, was selected for targeting the genes for the tobacco mosaic virus (TMV) coat protein (CP) and TMV RNA-dependent RNA polymerase (RdRP) to form HACC-dsRNA NPs. These NP-based dsRNAs were delivered to the plants using four different methods, including infiltration, spraying, root soaking, and pollen internalization. All four methods were able to reduce the seed-carrying rate of offspring seeds of the TMV-infected plants, with pollen internalization being the most effective in reducing the TMV-carrying rate from 95.1 to 61.1% in the control group. By measuring the plant uptake of fluorescence-labeled NPs and dsRNAs, the transportation of the HACC-dsRNA NPs into the plants was observed, and the uptake of dsRNA in combination with small RNA sequencing was further confirmed, resulting in the silencing of homologous RNA molecules during the topical application. The results demonstrated that the incidence of TMV infection was reduced by various degrees via RNAi induction without the need to develop transgenic plants. These results demonstrate the advantages of NP-based RNAi technology in breeding for disease resistance and developing a new strategy for virus-resistant breeding in plants.

Background Lysine 2-hydroxyisobutyrylation (Khib) is a novel and conserved post-translational modification (PTM). Frankliniella occidentalis are economically important agricultural pests globally and also notorious for vectoring destructive plant viruses. To better study the disease transmission mechanism of F. occidentalis , it is necessary to conduct in-depth analysis of it. So far, no Khib modification of insects has been reported. Results In this study, a proteome-wide analysis of Khib modifications in F. occidentalis was analyzed for the first time through the combination of high performance liquid chromatography fractionation technology and 2-hydroxyisobutyrylated peptide enrichment and other advanced technologies, 4093 Khib sites were identified on 1125 modified proteins. Bioinformatics and functional enrichment analyses showed that Khib-modified proteins were significantly enriched in many cell compartments and pathways, especially related to various cellular components and biological processes, and were more concentrated in ribosomes and proteasome subunits, involved in energy metabolism, protein synthesis and degradation, compared to the other nine species including Japonica rice, Homo sapiens , P. patens , Botrytis , Ustilaginoidea virens , Saccharomyces cerevisiae , T. gondii , C. albicans , and F. oxysporum . And Khib sites on virus-interacting insect proteins were discovered for the first time, such as cyclophilin and endoCP-GN. Conclusions After three repeated experiments, we found a total of 4093 Khib sites on 1125 proteins. These modified proteins are mainly concentrated in ribosomes and proteasome subunits, and are widely involved in a variety of critical biological activities and metabolic processes of F. occidentalis . In addition, for the first time, Khib modification sites are found on the proteome of F. occidentalis , and these sites could be acted as for the virus interaction, including cyclophilin and endoCP-GN. The global map of 2-hydroxyisobutyrylation in thrips is an invaluable resource to better understand the biological processes of thrips and provide new means for disease control and mitigation of pest damage to crops.