Banana wilt disease jeopardizes global banana production, exacerbated by the overuse of fungicides. The large-scale application of fungicides in soil provides certain preventive effects, but achieving precise and target control at the infection site during disease outbreaks remains challenging. To address these issues, an innovative amino acid-based nano-delivery system is constructed to deliver the non-systemic fungicide fludioxonil (Flu) to banana rhizomes. This system utilizes tryptophan (Trp), which can be mediated by amino acid transporters, and polysuccinimide (PSI) to load Flu through multiple mechanisms, enabling precise targeted delivery. By employing hydrogen bonding and π-π stacking interactions, the Flu-PSI-Trp17 achieves a loading efficiency of 53.74%, which enhances leaf deposition and causes severe damage to the mycelial structure of Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc TR4). Notably, targeted transport allows nanoparticles to accumulate in banana rhizomes, significantly mitigates disease-induced metabolic defects and improves the protective efficacy against Fusarium wilt in potted experiments by approximately 5-fold. Furthermore, the system demonstrates excellent biocompatibility and low phytotoxicity. Overall, this work provides a feasible approach for constructing targeted nano-delivery systems for agrochemicals, highlights the importance of developing amino acid-based nanocarriers with targeting capability, and offers a promising solution for advancing precise agriculture.

Root-knot nematodes pose a significant threat to global tomato production. Although avermectin (AVM) is an effective nematicide, its application is limited by difficulties in absorption and transportation in plants, poor photostability, short duration of action, and the complex preparation process of its emulsifiable concentrate (EC) formulations. This study presented a novel carrier-free nanoparticles (NPs) of AVM and phenylalanine (Phe) prepared through a simple self-assembly method. The resulting AVM-Phe NPs were comprehensively characterized, confirming the successful self-assembly of AVM and Phe via hydrogen bonding and hydrophobic interactions. The results indicated that AVM-Phe NPs exhibited a 30.77% enhancement in UV resistance and a 16.67% improvement in leaf retention capacity. In vitro studies demonstrated that AVM-Phe NPs can be ingested by nematodes and alter the mode of action of AVM, resulting in a significant increase of 8.15 times compared to AVM Technical grade in nematicidal efficacy. Furthermore, AVM-Phe NPs facilitated the transportation of AVM in tomato seedlings with a transportation rate of 3 times compared to AVM NPs, and there was a notable accumulation in the vascular bundles after the foliar application, which are the site of nematode infestation. This research offers a green, efficient, and innovative strategy for the application of biopesticides, potentially mitigating the current challenges associated with AVM use.

Succinate dehydrogenase inhibitors (SDHIs) have been widely employed as agricultural fungicides due to their effectiveness and low toxicity. Recently, sulfonyl fluoride (R-OSO2F) has emerged as a promising active moiety in pesticide development. Previously, our team identified aryl sulfonyl fluoride compounds with strong antifungal activity. Expanding on this, we incorporated the sulfonyl fluoride group into the heterocyclic amide scaffold of SDHIs, designing and synthesizing 56 novel aniline-based sulfonyl fluoride derivatives. Their structures were confirmed using 1H NMR, 13C NMR, and high-resolution mass spectrometry (HRMS). Antifungal activity assays showed that compound 4a exhibited the highest inhibitory activity against Rhizoctonia solani (EC50 = 2.89 μg/mL). In bioassays on rice leaves and pot experiments, compound 4a demonstrated 50.75% protective efficacy and 32.43% curative efficacy at 200 μg/mL, comparable to that of commercial SDHI fungicide boscalid. Enzyme inhibition assays confirmed its potent suppression of succinate dehydrogenase (SDH), and molecular docking studies demonstrated binding modes similar to boscalid, with improved binding affinity attributed to the sulfonyl fluoride moiety. We developed a novel sulfonyl fluoride series, identifying 4a as a potent antifungal lead with SDHI-like action. These findings provide valuable insights and new directions for the development of sulfonyl fluorides as next-generation fungicides.

Currently, the control of phytopathogenic fungi faces challenges arising from chemical resistance and biosafety concerns. Triazole fungicides (TFs), as ergosterol biosynthesis inhibitors, possess broad‑spectrum and high‑efficacy characteristics. However, their single target site has led to increasing field resistance, highlighting the urgent need to discover molecular targets with novel mechanisms of action. Although P‑type ATPases drive fungal pathogenicity, their potential as pesticide targets remains unexplored. Here, we identified APT5 as a broad-spectrum target protein commonly recognized by multiple TFs and elucidated its interaction mechanism. Using Magnaporthe oryzae (M. oryzae) as a model, in vivo experiments revealed that knockout of MoAPT5 inhibited mycelial growth and reduced the sensitivity of M. oryzae to TFs, whereas complementation restored both growth and sensitivity. Further pathogenicity assays showed that the ΔMoapt5 mutant exhibited reduced conidial production, abnormal conidial morphology, delayed germ tube elongation, and decreased pathogenicity on rice leaves, as well as markedly reduced sensitivity to the protective and curative effects of TFs. In vitro binding assays demonstrated that TFs exhibited strong affinity for MoAPT5 and inhibited its ATPase activity by approximately 50%. Molecular docking and dynamics simulations further revealed that TFs bind primarily to a hydrophobic pocket formed by transmembrane helices TM2, TM4, TM5, and TM6 of MoAPT5, with the complex remaining conformationally stable. These findings provide new insights into the multi-target mechanism of TFs and offer a potential target for overcoming existing resistance issues and developing novel fungicides.

ABSTRACT Clarifying the intrinsic impact of nanosizing on pesticide biointeractions remains challenging because most nanopesticides are formulated with nanocarriers or surfactants that may influence these interactions. Here, we report a disaggregation strategy for fabricating nanopesticides without nanocarriers or surfactants. Adding acetic acid to an emamectin benzoate (EB) suspension yielded a stable EB nanopesticide (HOAc‐EB) with an average particle diameter of 7 nm. In HOAc‐EB, acetic acid disrupted hydrogen bonding in EB rather than forming complexes with it. Nanosizing significantly increased bioactivity against Megalurothrips usitatus and Meloidogyne enterolobii , improved foliar and root penetration, and reduced soil adsorption. Compared with EB, HOAc‐EB reduced the 24‐h LC 50 by 91% against Megalurothrips usitatus (3.02 to 0.280 mg·L −1 ) and by 56% against Meloidogyne enterolobii (18.5 to 8.1 mg·L −1 ), along with 5.1‐fold higher peak foliar penetration in cowpea and 4.4‐fold higher peak root penetration in chili pepper at 1000 mg·L −1 . Field trials confirmed improved control of Meloidogyne enterolobii . HOAc‐EB was safe for crops at 1000 mg·L −1 and showed no greater toxicity than EB in zebrafish, earthworms, or mice. This study provides insights into the impact of nanosizing on pesticide biointeractions. The simple, eco‐friendly, and cost‐effective construction approach advances the practical production and agricultural applications of HOAc‐EB.

ABSTRACT In agriculture, propiconazole (PCZ) controls excessive growth in flowering Chinese cabbage but poses dietary safety risks due to residue accumulation. Therefore, identifying novel PCZ targets and breeding PCZ-free cultivars is critical for the safe production of flowering Chinese cabbage. Here, we identified three P4-ATPase flippase homologs aminophospholipid ATPase 3 (BraALA3a/b/c) in flowering Chinese cabbage that function as sensitive targets for PCZ. These proteins exhibit high binding affinity for PCZ, which directly inhibits their ATPase activity. Overexpression of the BraALA3 homologs enhanced plant growth and increased sensitivity to PCZ, whereas knockdown led to dwarfism and reduced sensitivity. Based on these findings, we identified editable active sites via protoplast-based screening. Genetic transformation of one such site yielded BraALA3a/ braala3a K200T mutant lines, which displayed a dwarf and compact architecture. These findings provide a precise molecular target for developing PCZ-free germplasm in flowering Chinese cabbage through gene editing.

Isoxazolines, which act as modulators of the GABA receptor (GABAR), are significant agrochemicals for controlling of resistant pest populations. However, their sustainable application is often hindered by toxicity to beneficial non-target organisms, particularly pollinators. To investigate the potential for reducing the toxicity of isoxazoline analogs to honeybees, a series of novel isoxazoline-thiazole derivatives were designed and synthesized using the "ring-closure" strategy. Bioassays demonstrated that these compounds retained promising insecticidal activity. Among them, compounds 9m and 9l exhibited excellent insecticidal activity against Spodoptera litura, with LC50 values of 0.87 and 0.80 mg/L, respectively, comparable to that of fluxametamide (0.65 mg/L). Moreover, compound 9m also showed good insecticidal activity against Plutella xylostella. Importantly, compound 9m displayed reduced acute toxicity to honeybees, with an LD50 of 8.98 μg/bee (indicating moderate toxicity), whereas fluxametamide has an LD50 of 1.24 μg/bee (indicating high toxicity). Integrated molecular docking, MD simulations, and DFT calculations suggested that the incorporation of the thiazole moiety contributed to favorable target binding while diminishing interactions with honeybee GABAR. In conclusion, these isoxazoline-thiazole derivatives may serve as potential candidates and lead structures for the development of novel insecticides with low toxicity to honeybees.

ObjectiveTo isolate and identify the pathogen responsible for Pineapple (Ananas comosu) fruitlet core rot (FCR), and screen control agents and field-based combination treatments, thereby providing a basis for studying the disease pattern and mechanism as well as green prevention and control.MethodPathogens were isolated and purified, and the pathogenicity was verified using Koch’s law. The species of pathogens were determined according to the morphological characteristics of the strains and multi-gene sequence analysis. The inhibitory effects of the pathogen on five single agents and eight compound agents were determined using the mycelial growth rate method. ResultNine genera of fungal strains were obtained by isolation and purification, among which two strains could infect pineapple fruitlets and cause brown spots. Combining morphological characterization with multi-gene sequence analysis, the pathogenic fungi that caused FCR were identified as Fusarium verticillioides LDL-3 and Talaromyces funiculosus LL-3, with LDL-3 being more pathogenic than LL-3. The results of fungicide virulence tests showed that flucytosine, feniconazole, pyraclostrobin, imidacloprid manganese salts and compound formulations had effects on the growth of the pathogens. Among these agents, imidacloprid manganese salts had a better inhibitory effect on LDL-3 with an EC50 of 0.009 μg/mL, while pyraclostrobin showed a better inhibitory effect on LL-3 with an EC50 of 0.134 μg/mL. Among eight compound formulations, benzyl · pyraclostrobin (volume ratio of difenoconazole and pyraclostrobin was 3∶1) inhibited both pathogens effectively with EC50 less than 0.2 μg/mL. The combined use of insecticide, fungicide and growth regulator could effectively inhibit the pathogen growth with EC50 less than 0.2 μg/mL. ConclusionThe FCR is mainly caused by Fusarium sp. and Talaromyces sp.. Benzyl · pyraclostrobin (3∶1, v/v) effectively inhibits these pathogens and offers a simplified and efficient solution for field control during flower induction and fruit expansion.

The ecological and health risks posed by pesticides entering soil environments are a growing concern. As a new GABA receptor antagonist insecticide, the environmental fate, ecotoxicity and potential transformation products (TPs) of pyraquinil in soil have remained ambiguous. In this study, degradation kinetics, pathways, mechanisms, and toxicities of pyraquinil in five representative soils under aerobic/anaerobic/sterile conditions were investigated for the first time. Pyraquinil exhibited a half-life (DT50) of 25.5-79.3 days, with degradation accelerated by elevated soil pH, OMC, CEC, and microbial abundance. Using suspect and nontarget screening strategies, 33 TPs generated in soil were identified via UHPLC-Orbitrap-HRMS, with the fused heterocyclic scaffold remaining stable across all TPs. Among these, 23 TPs were newly reported, most exhibiting significant structural divergence from pyraquinil. Degradation pathways varied by soil type and oxygen availability, primarily involving oxidation, hydrolysis, decarboxylation, hydroxylation, and dehydration. Density functional theory calculations demonstrated the deprotonated form of pyraquinil is more reactive than its protonated counterpart, and revealed that these degradation reactions are kinetically and thermodynamically feasible. Experimental and ECOSAR-predicted data indicated low acute toxicity of pyraquinil to earthworms (LC50 > 100 mg/kg), whereas nearly 30% of the TPs exhibited higher toxicities to aquatic organisms than the parent compound. These results systematically elucidate pyraquinil's environmental fate and highlight the ecological risks posed by TPs, providing critical insights for risk assessment of pyraquinil in soil.

The widespread use and persistence of quaternary ammonium compounds (QACs) in food environments raise concerns about their detection at trace levels in complex matrices. In this study, we developed a negatively charged 4-mercaptobenzenesulfonic acid functionalized SiO2@Au core-shell nanocomposite (MBSA-SiO2@Au) that serves as both a selective adsorbent and an efficient surface-assisted laser desorption/ionization (SALDI) matrix. The sulfonate modified gold shell provides a stable negative surface potential to promote electrostatic interactions with cationic QACs, and their plasmonic properties synergistically enhance laser energy absorption and ionization efficiency. Critical enrichment parameters, including adsorbent amount, solution pH, adsorption time, and redispersion volume, were optimized. The optimized method achieved limits of detection of 0.5 ng/mL for paraquat (PQ), dodecyltrimethylammonium chloride (DTAC), and didodecyldimethylammonium chloride (DDAC), and 1.5 ng/mL for dodecyldimethylbenzylammonium chloride (DBAC), with excellent linearity (R2 ≥ 0.9843), achieving an enrichment factor of approximately 33.3, and a 20- to 40-fold sensitivity enhancement after enrichment. Application to real food samples, including white radish, carrot, and potato, revealed no detectable QAC residues and satisfactory recoveries ranging from 87.0 to 108.8%. The bifunctional MBSA-SiO2@Au-based SALDI platform minimizes organic solvent consumption and offers a rapid, green, and sensitive approach for monitoring QACs in food safety applications.

Nanopesticides offer unique advantages for the targeted delivery of agrochemicals against destructive plant vascular diseases due to their superior in planta transport capability. However, a fundamental understanding of how nanopesticides penetrate crop tissues and undergo in vivo translocation remains elusive. This knowledge gap severely limits the rational design of targeted nanopesticides and contributes to pesticide waste and ecological contamination. Here, a serine-functionalized polysuccinimide nanocarrier (PSI-Ser) was developed, onto which fludioxonil (Flu) was loaded, yielding Flu-PSI-Ser nanoparticles (NPs) with an average diameter of approximately 80 nm. By comparing the uptake and visualization of Flu-PSI-Ser NPs before and after abscisic acid (ABA) treatment, the nanopesticide was found to the leaf epidermis through stomata. After entering the leaf, the NPs were primarily distributed in the extracellular space, with almost no internalization into the mesophyll cells. During the apoplastic transport of Flu-PSI-Ser NPs, a small fraction of Flu molecules released from the NPs can enter the cytoplasm. Notably, Flu-PSI-Ser NPs demonstrated efficient phloem translocation, allowing systemic movement from treated leaves to underground rhizomes. As a non-systemic fungicide, Flu-PSI-Ser NPs enabled Flu accumulation in rhizome tissues up to 0.601 mg/kg. Owing to their optimized biodistribution and vascular-targeted delivery, Flu-PSI-Ser NPs exhibited approximately 3.2-fold higher control efficacy against banana Fusarium wilt compared with Flu SC concentrate. This study clarifies the tissue-level uptake and long-distance translocation pathways of amino acid-modified nanopesticides in crops and provides a rational delivery strategy for vascular disease management and the development of sustainable agriculture.

Currently, the control of phytopathogenic fungi faces challenges arising from chemical resistance and biosafety concerns. Triazole fungicides (TFs), as ergosterol biosynthesis inhibitors, possess broad-spectrum and high-efficacy characteristics. However, their single target site has led to increasing field resistance, highlighting the urgent need to discover molecular targets with novel mechanisms of action. Although P-type ATPases drive fungal pathogenicity, their potential as pesticide targets remains unexplored. Here, we identified MoAPT5 as a potential target protein commonly recognized by multiple TFs and elucidated its interaction mechanism. Using Magnaporthe oryzae (M. oryzae) as a model, in vivo experiments revealed that knockout of MoAPT5 inhibited mycelial growth and reduced the sensitivity of M. oryzae to TFs, whereas complementation restored both growth and sensitivity. Further pathogenicity assays showed that the ΔMoapt5 mutant exhibited reduced conidial production, abnormal conidial morphology, delayed germ tube elongation, and decreased pathogenicity on rice leaves, as well as markedly reduced sensitivity to the protective and curative effects of TFs. In vitro binding assays demonstrated that TFs exhibited strong affinity for MoAPT5 and inhibited its ATPase activity by approximately 50%. Molecular docking and dynamics simulations further revealed that TFs bind primarily to a hydrophobic pocket formed by transmembrane helices TM2, TM4, TM5, and TM6 of MoAPT5, with the complex remaining conformationally stable. These findings provide new insights into the multi-target mechanism of TFs and offer a potential target for overcoming existing resistance issues and developing novel fungicides.

Abstract Propiconazole (PCZ) is widely misused growth regulator in leafy Brassica vegetables. Developing green strategies for managing plant architecture has become an urgent agricultural priority. Here, we identified from a membrane-protein-defective yeast library a P4-ATP phospholipid flippase, aminophospholipid ATPase 3 (ALA3), as a target sensitive to PCZ. ALA3 exhibits high binding affinity for PCZ, which inhibits its ATPase activity. Knockdown of ALA3 rendered yeast, Arabidopsis , and Brassica rapa less sensitive to PCZ and conferred a growth-inhibited phenotype. This dwarfing phenotype is mediated through the interaction between ALA3 and CYP51G1 that jointly acts within the brassinosteroid regulatory pathway. Furthermore, we identified lead compounds A01 and A15 as ALA3-targeting agents, and compared to PCZ, they display superior binding affinity and reduced toxicity. Our work establishes ALA3 as a key mediator of PCZ-induced dwarfism and provides dual strategies—creating promising varieties through gene editing and developing targeted green pesticides—to reduce PCZ use. Teaser Targeting ALA3 reduces PCZ use through gene-edited varieties and green pesticides.

In contrast to bactericides, elicitors induce plant immune systems to defend against pathogen attack and avoid potential damage to the environment. However, the energy cost caused by the continuous activation of immunity leads to the inhibition of plant growth, which has limited the agricultural application of a large number of elicitors. Here, we identified a natural elicitor 3,4-dihydroxy-3-methyl-2-pentanone (DMPN) that can induce disease resistance in plants. DMPN contains four stereoisomers (3R,4S), (3S,4R), (3R,4R) and (3S,4S), which exhibit different induced resistance activities in Arabidopsis thaliana but do not inhibit plant growth. B1 is different from the other three isomers in that it only induces disease resistance to the necrotrophic pathogen Erwinia carotovora instead of the biotrophic pathogen Pseudomonas syringae, and the remaining isomers is effective for both pathogens. When it comes to threo-isomers B1 (3R,4S) and B2 (3S,4R), transcriptomic and gene expression analysis reveal that both B1 and B2 activated the jasmonic acid (JA)/ethylene (ET) and chitin-mediated signalling pathways. B2 also activated the salicylic acid (SA) pathway and upregulated a wider range of defence-related genes. These findings indicate that stereoconfiguration critically influences elicitor bioactivity. In summary, we reported a natural stereoisomeric elicitor, DMPN, which can elicit the plant defence response in Arabidopsis thaliana without inhibiting plant growth and revealed the differential inducing effects on the plant immune system of its four isomers.

The development of novel phenylpyrazole insecticides is essential for managing pest resistance. Nicofluprole, a phenylpyrazole insecticide, serves as a valuable lead compound for structural optimization due to its insecticidal activity. In this study, a series of phenylpyrazole derivatives was designed and synthesized via an isosteric ring exchange strategy, in which the 2-chloropyridine ring of nicofluprole was replaced with substituted phenyl rings. The corresponding compounds exhibited good insecticidal activity against Plutella xylostella, and compounds 9q and 9r showed LC50 values of 0.49 and 0.20 mg/L, respectively. Structure-activity relationship (SAR) analysis indicated that electron-withdrawing substituents on the phenyl ring were associated with enhanced insecticidal activity. Density functional theory (DFT) calculations and molecular docking studies were conducted to investigate the electronic properties and potential binding modes of representative compounds. The results provide mechanistic insight into how ring replacement and phenyl substitution modulate insecticidal activity within the phenylpyrazole scaffold.

As per- and polyfluoroalkyl substances (PFASs) are persistent pollutants with high bioaccumulation potential, insights into their absorption, transport, and spatial distribution in whole plants are critical for evaluating their ecological impact and risks. Here, we develop a novel gold nanoparticle-coated urea-linked organic polymer-functionalized TiO2 nanotube substrate for whole-plant imprinting mass spectrometry imaging (MSI) in negative-ion modes. We investigated the spatially and temporally distinctive absorption, translocation, and accumulation patterns of three PFASs with different carbon chain lengths (CCLs), including perfluorobutanesulfonate (PFBS, C4), perfluorohexanesulfonate (PFHxS, C6), and perfluorooctanesulfonate (PFOS, C8), in whole hydroponically grown cowpea plants. The MSI results reveal that PFBS with the shortest CCLs presents the highest mobility that is distributed throughout the entire plant within 6 h, whereas PFOS presents limited transport and accumulates primarily in the roots and stems, even after prolonged exposure. Cross-sectional MSI further reveals that both PFBS and PFHxS easily penetrate root tissues and accumulate in the vascular cylinder, whereas PFOS is likely restricted by the Casparian strip in the endodermis, rendering it hard to enter the phloem and xylem of roots. Collectively, this whole-plant imprinting MSI serves as a powerful tool for deciphering the CCL-dependent translocation mechanisms of PFASs in plants, contributing to understanding and pollution management of PFASs in an ecological environment.

The evolutionary arms race between insectivorous bats and their insect prey is a classic paradigm of acoustic predation and evasion, with insects having evolved sophisticated auditory countermeasures. Both bats and insects also rely heavily on olfaction for key behaviors, such as social communication. Moreover, predator-derived odors are well-established as risk cues in many other predator–prey systems. However, whether olfaction plays a role in the bat–insect arms race remains unknown. Here, we unveil a previously unknown olfactory dimension to this interaction. We demonstrated that the body odor of the insectivorous bat Scotophilus kuhlii triggered robust avoidance and electrophysiological antennal responses in a common cricket prey, Loxoblemmus equestris. We identified limonene as a behaviorally active volatile in bat odor that elicited electrophysiological responses in cricket antennae and was sufficient to elicit avoidance in crickets. Field experiments confirmed that limonene exposure reduced cricket calling activity, demonstrating the ecological relevance of this cue. Our findings establish that insects can detect and initiate avoidance of phylogenetically distant vertebrate predators via olfaction, a process that could be mediated by the elemental perception of individual odor compounds. This work broadens the sensory framework of a classic predator–prey system, and highlights olfactory eavesdropping as a functional strategy in phylogenetically distant predator–prey systems.

Fluralaner, as a broad-spectrum and highly effective pesticidal molecule, is severely limited from agricultural application due to its high toxicity to honeybees. Herein, novel pro-pesticides were prepared by covalently linking fluralaner to polyethylene glycol and long-chain fatty acids, respectively, and were subsequently self-assembled into regular nano-micelles to reduce the toxicity of fluralaner to honeybees. The results showed that the obtained pro-pesticides self-assembled into nano-micelles without using any adjuvants, and the prepared nano-micelles showed spherical morphology, a low polydispersity index, strong negative charges, good surface activity and excellent maximum retention on hydrophobic leaves because of the amphiphilic structure. The toxicity of fluralaner prodrug molecules coupled with different fragments against Apis mellifera was more than 50 times lower than that of fluralaner. The solubility of compound E1 was increased by 68.4 times and was systemically distributed in all parts of Chinese flowering cabbage (Brassica rapa var. parachinensis; abbreviation: CS) seedlings. Therefore, the self-assembly nanotechnology of this prodrug conjugate is expected to improve the effective utilization rate of pesticides and reduce the toxicity of pesticides to the ecology and environment.