Pesticide Biochemistry and Physiology最新文献

{"title":"Premature termination codon mutations in osmosensor-like histidine kinase FgOs1 endow Fusarium graminearum with fludioxonil resistance","authors":"Yaru He ,&nbsp;Zikuo Li ,&nbsp;Ziyun He ,&nbsp;Aoran Wang ,&nbsp;Xuheng Gao ,&nbsp;Haoxue Xia ,&nbsp;Shanyue Zhou ,&nbsp;Wende Liu ,&nbsp;Guangfei Tang","doi":"10.1016/j.pestbp.2025.106633","DOIUrl":"10.1016/j.pestbp.2025.106633","url":null,"abstract":"<div><div><em>Fusarium graminearum</em> is a devastating disease in cereal production, causing a loss in grain production. Fludioxonil, a phenylpyrrole fungicide, has been registered for disease management of many crops. However, the resistance mechanism of <em>F. graminearum</em> to fludioxonil has not been systematically analyzed. This study elucidates the molecular basis of fludioxonil resistance in <em>F. graminearum</em> by demonstrating that premature termination codon mutations in the osmosensor-like histidine kinase gene <em>FgOs1</em> are causative factors. Two independent mutations were identified in fludioxonil-resistant mutants, leading to truncated <em>FgOs1</em> proteins: a FgOs1^Q140STOP mutation locating at N-terminal and an FgOs1^R1183STOP mutation locating at the REC signaling domain. Homologous gene replacement and complementation assays confirmed that these mutations specifically confer high-level resistance to fludioxonil (resistance factor &gt; 1000) without cross-resistance to tebuconazole, phenamacril, or carbendazim. Notably, <em>FgOs1</em>-mutated strains exhibited heightened sensitivity to osmotic and metal ion stresses, suggesting that the premature termination codon-induced protein truncation impairs osmoregulation and ion homeostasis pathways. Phylogenetic analysis revealed that the mutated residues are highly conserved across diverse fungal species, underscoring their functional importance. These findings uncover a previously unrecognized role of <em>FgOs1</em> in antifungal resistance and provide critical targets for developing innovative strategies to manage fludioxonil resistance in <em>F. graminearum</em>.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"215 ","pages":"Article 106633"},"PeriodicalIF":4.0,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144906991","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Neonicotinoids and human health: Environmental fate, toxicity mechanisms, and future directions","authors":"Kangming Ji ,&nbsp;Yu Rong ,&nbsp;Zihang Zhang ,&nbsp;Yunjie Chen ,&nbsp;Ruixin Hu ,&nbsp;Yuchen Chu ,&nbsp;Yuyi Tang ,&nbsp;Lijun Qian ,&nbsp;Chuang Yang","doi":"10.1016/j.pestbp.2025.106636","DOIUrl":"10.1016/j.pestbp.2025.106636","url":null,"abstract":"<div><div>Neonicotinoid insecticides are among the most widely used agricultural chemicals globally, playing a significant role in controlling pests. However, their unintended effects on non-target species, particularly humans, have sparked global health and environmental concerns. This article reviews the environmental distribution, metabolic pathways, and mechanisms of action of neonicotinoids, as well as their toxicity to multiple human systems, including the digestive, respiratory, nervous, and endocrine systems. We dissect the molecular pathways linking neonicotinoid exposure to oxidative stress and apoptotic cell death, and discuss other potential harms, such as nephrotoxicity, cardiotoxicity, reproductive toxicity, pancreatic toxicity, and immunotoxicity. Additionally, we emphasize the importance of monitoring neonicotinoids and their metabolites and propose key areas for future research, including in-depth studies of toxicity mechanisms, assessments of combined toxicity effects, the development of biomarkers, and strategies to reduce environmental release and human exposure. The conclusion of this article underscores the necessity of taking multifaceted measures to reduce the environmental release and human exposure of neonicotinoids, as well as the urgency of conducting interdisciplinary research to fully understand the environmental fate and health impacts of neonicotinoids. Through these efforts, we can better protect human health from the potential hazards of neonicotinoids and promote the sustainable use of pesticides.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"215 ","pages":"Article 106636"},"PeriodicalIF":4.0,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144890326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Structural plasticity underlies the binding capacities and specificities of chemosensory proteins towards various pesticides","authors":"Jinbing Zhang,&nbsp;Qian Jia,&nbsp;Wei Xie","doi":"10.1016/j.pestbp.2025.106647","DOIUrl":"10.1016/j.pestbp.2025.106647","url":null,"abstract":"<div><div>Chemosensory proteins (CSPs) are small proteins that play important roles in insect physiology. <em>Spodoptera litura</em>, a notorious pest, causes considerable losses to the agricultural industry. Recent studies have linked the pesticide resistance of <em>S. litura</em> to CSPs, but the resistance mechanism remains poorly understood due to a dearth of biochemical studies and structural information on the CSP-ligand complexes. In this study, we report the crystal structure of <em>Spodoptera litura</em> CSP3 (SlCSP3). A comparative structural analysis of apo-SlCSP3 with its ligand-bound counterparts or AlphaFold2-predicted CSPs reveals a previously overlooked conformational state with an expanded pocket size. Furthermore, molecular dynamics (MD) simulations highlight the inherent structural plasticity of CSPs. Through virtual screening of potential CSP ligands, numerous agrochemical candidates were identified, with pyrethroids emerging as the primary category. Following docking studies demonstrated that the structurally diverse C-termini of CSPs dictate their binding specificities and affinities. This study offers insights into the structural underpinnings of differential binding propensities and specificities of CSPs, thereby shedding light on their pertinent mechanisms of drug resistance.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"215 ","pages":"Article 106647"},"PeriodicalIF":4.0,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144890328","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fipronil transformation in aquatic ecosystems and ecotoxic effects on fish populations","authors":"Surya Prakash Pandey ,&nbsp;Maryam Fatma ,&nbsp;Rakesh Bhaskar ,&nbsp;Sung Soo Han","doi":"10.1016/j.pestbp.2025.106641","DOIUrl":"10.1016/j.pestbp.2025.106641","url":null,"abstract":"<div><div>The phenylpyrazole insecticide fipronil (FIP) is a wide-range agrochemical that is extensively used in agricultural, veterinary, and household settings. It is oxidized, reduced, hydrolyzed, and/or photolyzed in water into -sulfone, −sulfide, −amide, −desulfinyl, or fipronil-(13C3, cyano-13C) metabolites having varying degrees of ecotoxicity. Temperature, pH, organic matter content, and other environmental factors influence its bioavailability and toxicity in water and sediments. Excessive applications have raised concerns about FIP's ecotoxicological consequences on aquatic non-target organisms. This article discusses the impact of FIP's biotransformation on non-target invertebrates and vertebrates in aquatic systems. Being a gamma-aminobutyric acid (GABA) inhibitor, it is an efficient disruptor of the central nervous system of target insects and non-target aquatic species (both invertebrates and vertebrates), including fish. Widely distributed hormone receptors and neurotransmitters (including acetylcholine, dopamine, and serotonin) in the central hypothalamic and peripheral endocrine systems of fish are sensitive to FIP and its metabolites. Multibiomarker studies have demonstrated numerous sublethal effects on various physiological responses like endocrine disruption, oxidative stress, growth, and reproductive impairment. These effects lead to alterations in behavioral responses, feeding efficiency, and spawning success, ultimately lowering the long-term viability of fish populations. This review also comprehends the potential risks of FIP exposures to fish populations by emphasizing the susceptibility, physiological impairments, and risk assessment. Bioaccumulation studies indicate FIP-accumulation in freshwater and marine food webs, posing direct/indirect risk to piscivores of higher trophic levels. Further studies must elucidate mechanisms of sub-lethal effects, long-term bioaccumulation, and chemical exposure-ecosystem interaction to enhance the understanding of population-level effects.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"215 ","pages":"Article 106641"},"PeriodicalIF":4.0,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144903442","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phenotype MicroArray™ reveals the effects of azoxystrobin on the phenotypic characteristics of Rhizoctonia solani","authors":"Ligang Xiang ,&nbsp;Hancheng Wang ,&nbsp;Liuti Cai ,&nbsp;Lili Chen ,&nbsp;Jianyu Meng ,&nbsp;Mingxia Wen ,&nbsp;Tom Hsiang ,&nbsp;Wenjian Zhang","doi":"10.1016/j.pestbp.2025.106637","DOIUrl":"10.1016/j.pestbp.2025.106637","url":null,"abstract":"<div><div><em>Rhizoctonia solani</em> is a soil-borne pathogen that poses a significant threat to agricultural productivity. Azoxystrobin, a potent fungicide, is extensively utilized for the prevention and control of diseases caused by <em>R. solani</em>. This study evaluated the effects of azoxystrobin (1, 10, and 100 μg/mL) on the substrate utilization patterns and phenotypic characteristics of <em>R. solani</em> using the Phenotype MicroArray™ system. Without azoxystrobin exposure, <em>R. solani</em> demonstrated remarkable metabolic capacity and robust environmental adaptability, utilizing 97.89 % of carbon sources, 98.94 % of nitrogen sources, and 100 % of phosphorus, sulfur, nutrient supplement, and peptide nitrogen sources, growing under 80 (83.33 %) osmotic pressures and 91 (94.79 %) pH conditions. The effect of azoxystrobin on the metabolic capacity of <em>R. solani</em> exhibited a nonlinear correlation with its concentration. At concentrations of 1, 10, and 100 μg/mL, azoxystrobin induced a loss of phenotype in 699, 855, and 862 substrates, and a gain of phenotype in 229, 74, and 68 substrates. The key downregulated KEGG pathways included metabolic pathways, ABC transporters, biosynthesis of secondary metabolites, starch and sucrose metabolism, purine metabolism, and biosynthesis of amino acids, among others. Notably, arbutin and salicin showed significant enhanced metabolic capacity under three azoxystrobin concentrations, involved in glycolysis/gluconeogenesis and phosphotransferase system (PTS), suggesting their role in fungicide response. These findings provide better insights into the adaptability of <em>R. solani</em> and the mechanisms of action of azoxystrobin.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"215 ","pages":"Article 106637"},"PeriodicalIF":4.0,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144890402","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Antifungal activity and action mechanism of thymol against rice blast fungus Magnaporthe oryzae","authors":"Huan Liu ,&nbsp;Ziyi Wang ,&nbsp;Guanfei Yu ,&nbsp;Chenxi Yan ,&nbsp;Yiming Liu ,&nbsp;Shengming Liu ,&nbsp;Shuzhen Deng","doi":"10.1016/j.pestbp.2025.106646","DOIUrl":"10.1016/j.pestbp.2025.106646","url":null,"abstract":"<div><div>Rice blast (<em>Magnaporthe oryzae</em>) is one of the most devastating disease in rice. There is an urgent need to find effective and environmentally friendly chemicals for the prevention and control of rice blast. In this study, we evaluated the antifungal activities of five plant-derived essential oils: black pepper essential oil, perilla essential oil, thyme essential oil, peony essential oil, and rosemary essential oil, against <em>M. oryzae</em>. Specifically, thyme essential oil exhibited an excellent antifungal activity at a concentration of 1 μL/mL. To ascertain the active ingredient, the oil was subsequently analyzed using gas chromatography–mass spectrometry (GC–MS). Thymol was identified as the primary active component and antifungal assays showed that thymol effectively inhibited spore germination and appressorium formation of <em>M. oryzae.</em> In addition, treatment of thymol impaired mycelial micromorphology and appressorium-mediated penetration. Biosafety assays indicated that thymol exhibited no phytotoxicity towards rice. Furthermore, transcriptome analysis indicated that thymol might significantly affect melanin biosynthesis pathway of <em>M. oryzae</em>. This inference was further supported by preliminary molecular docking analysis of thymol with four proteins (Hnr1, Lac8, Buf1, and Rsy1) involved in melanin biosynthesis pathway. Our study provides new insights into the potential mechanism of thymol against <em>M. oryzae</em>.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"215 ","pages":"Article 106646"},"PeriodicalIF":4.0,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144906988","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"FgSPF1 regulates DON biosynthesis and fungicide sensitivity in fusarium graminearum by regulating toxisome formation and membrane permeability","authors":"Furong Chen,&nbsp;Yunkai Gu,&nbsp;Chengxiang Dai,&nbsp;Yiping Hou","doi":"10.1016/j.pestbp.2025.106643","DOIUrl":"10.1016/j.pestbp.2025.106643","url":null,"abstract":"<div><div><em>Fusarium graminearum,</em> which causes <em>Fusarium</em> head blight (FHB), reduces crop yield and compromises wheat quality by producing mycotoxins such as deoxynivalenol (DON)<em>.</em> While SPF1 is well studied for preserving ER membrane integrity and ensuring transmembrane protein quality in yeast, its role in agriculturally important plant-pathogenic fungi like <em>F. graminearum</em> remains largely unexplored. Here, we demonstrate that FgSPF1, the ortholog of yeast SPF1 in <em>F. graminearum</em>, plays a pivotal role in regulating DON biosynthesis and fungicide sensitivity. ΔFgSPF1 mutants exhibited reduced virulence on wheat and significantly decreased DON production, which was directly attributed to the disruption of toxisome structure. Notably, ΔFgSPF1 mutants exhibited decreased sensitivity to seven types fungicides with distinct target site. Mechanistically, LC-MS analysis, conductivity measurements, and ultrastructural observations revealed that ΔFgSPF1 strains had enhanced cell membrane compactness and reduced membrane permeability, leading to decreased fungicide absorption and thus increased multi-fungicide resistance. Additionally, disruption of FgSPF1 impaired vegetative growth, reduced asexual sporulation, and abolished sexual reproduction due to actin polarity disruption and hyphal morphological changes. Collectively, this study identified a novel regulatory mechanism of FgSPF1 in pathogenicity and fungicides sensitivity of <em>F. graminearum</em>.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"215 ","pages":"Article 106643"},"PeriodicalIF":4.0,"publicationDate":"2025-08-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144890400","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selective interaction of BmNPV Bm65 with host proteins before and after ultraviolet radiation","authors":"Yutong Liu,&nbsp;Jiayin Huang,&nbsp;Guohui Li,&nbsp;Feifei Zhu,&nbsp;Qian Yu,&nbsp;Lindan Sun,&nbsp;Huiqing Chen,&nbsp;Liang Chen,&nbsp;Qi Ge,&nbsp;Shangshang Ma,&nbsp;Xiaoyong Liu,&nbsp;Keping Chen,&nbsp;Qi Tang","doi":"10.1016/j.pestbp.2025.106645","DOIUrl":"10.1016/j.pestbp.2025.106645","url":null,"abstract":"<div><div>Baculovirus biopesticides are highly susceptible to inactivation by ultraviolet (UV) radiation in sunlight. At present, the DNA repair mechanism in most baculoviruses after ultraviolet (UV) radiation is still unclear. Our previous research found that <em>Bombyx mori</em> nucleopolyhedrovirus Bm65 was a very important UV-specific endonuclease, and the knockout of Bm65 gene increased the sensitivity of BmNPV to UV radiation. Based on the important role of Bm65 in UV-induced DNA repair, we further explored Bm65-interacting proteins using immunoprecipitation and liquid chromatography-mass spectrometry (LC-MS) assays in this study. The interaction between host DNA repair proteins and Bm65 was detected only after UV radiation. Furthermore, when the viruses or hosts were separately damaged by UV radiation, the DNA repair proteins interacting with Bm65 were not completely consistent. We showed for the first time that BmRad23, a key DNA damage recognition protein of host was a UV-dependent interactor of Bm65, and interacted with Bm65 only after UV radiation. It was further found that Bm65 relied on the interaction with BmRad23 to accurately localize at the sites of UV-damaged viral DNA. These results suggested that Bm65 selectively interacted with host proteins before and after UV radiation, and there might be different pathways that Bm65 participated in the UV-induced DNA repair of viruses or hosts.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"214 ","pages":"Article 106645"},"PeriodicalIF":4.0,"publicationDate":"2025-08-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144865532","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metal–phenolic networks improve biocontrol effect of Bacillus velezensis DM14 against pear anthracnose","authors":"Jianfeng Du ,&nbsp;Heng Li ,&nbsp;Shenjie Liu ,&nbsp;Jiawei Tang ,&nbsp;Xin Zhao ,&nbsp;Yiming Liu ,&nbsp;Limin Wu ,&nbsp;Yue Li ,&nbsp;Qixiong Gao ,&nbsp;Fuxin Sun ,&nbsp;Ninghai Lu ,&nbsp;Xueliang Tian ,&nbsp;Yang Jiao ,&nbsp;Chuanjun Wang","doi":"10.1016/j.pestbp.2025.106638","DOIUrl":"10.1016/j.pestbp.2025.106638","url":null,"abstract":"<div><div>Pear anthracnose is a serious postharvest disease that affects pear production. The use of antagonistic microorganisms for biological control provides a promising alternative to fungicides. Unfortunately, these microorganisms are susceptible to harsh environmental conditions, which limits their field application. To overcome this limitation, we investigate the biocontrol potential of <em>Bacillus velezensis</em> DM14, which was isolated from the surface of healthy pear fruits and exhibits significant inhibitory effects on <em>Colletotrichum fructicola</em>. Microscopic analysis revealed that <em>C. fructicola</em> cells exposed to secreted fermentation extract raw fermentation extract (RFE) of DM14 underwent significant morphological changes, including intracellular disruptions such as cytoplasmic disintegration and vacuolization, ultimately leading to cell death. While the fermentation extract of DM14 was effective in vitro, enhancing the survival of live DM14 cells is essential for practical application. Therefore, we use a one-step protection based on metal-phenolic network (MPN) encapsulation for DM14. This MPN-coated DM14 has shown a higher colonization rate under oxidative stress conditions in fruit wounds. Importantly, compared with uncoated DM14, the MPN-coated DM14 enhanced the prevention of pear anthracnose by approximately 37 %. Furthermore, the MPN coating strategy improved the tolerance to oxidative microenvironments in infected fruit wounds. Overall, this microbial encapsulation strategy is a promising way to protect fragile antagonistic microorganisms, providing attractive avenues in sustainable agriculture.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"215 ","pages":"Article 106638"},"PeriodicalIF":4.0,"publicationDate":"2025-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144890403","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synergistic disruption of detoxification, immunity, and neural pathways in honeybees following co-exposure to cyantraniliprole and difenoconazole","authors":"Xuan Liu ,&nbsp;Wenchao Zhu ,&nbsp;Xiaojun Gai ,&nbsp;Shaoqing Liu ,&nbsp;Chunxiao Wang ,&nbsp;Lu Lv ,&nbsp;Yanhua Wang ,&nbsp;Xuejing Wang ,&nbsp;Zeqi Lu ,&nbsp;Zhixin Wang","doi":"10.1016/j.pestbp.2025.106640","DOIUrl":"10.1016/j.pestbp.2025.106640","url":null,"abstract":"<div><div>The diamide insecticide cyantraniliprole (CYA) and the triazole fungicide difenoconazole (DIF) are frequently co-detected in bee-related matrices. However, the interactive effects of these compounds on honey bee (<em>Apis mellifera</em> L.) physiology remain insufficiently elucidated. Our results revealed that co-exposure to CYA and DIF elicited a pronounced acute synergistic toxicity. Biochemical assays demonstrated significant elevations in malondialdehyde (MDA) level, superoxide dismutase (SOD), and caspase-3 (CASP-3) activities across all treatments, with the most marked alterations occurring under co-exposure conditions. These data pointed to exacerbated oxidative stress and mitochondrial impairment when both pesticides were present concurrently. At the transcriptional level, notable dysregulation was observed in genes associated with apoptosis (<em>caspase-1</em>), detoxification (<em>CYP4G11</em>), immune modulation (<em>dorsal-2</em>), and lifespan regulation [<em>vitellogenin</em> (<em>vtg</em>)]. Notably, co-exposure intensified gene expression changes beyond those induced by single-pesticide treatments, underscoring a compound interaction that amplified cellular stress responses. These findings demonstrated that both CYA and DIF, especially in combination, disrupted critical physiological pathways in honey bees, compromising their detoxification capacity, immune integrity, and longevity. These insights not only unraveled key mechanistic underpinnings of pesticide mixture toxicity but also emphasized the urgent need for regulatory frameworks that address the mixture risks posed by agrochemical co-exposures in pollinator populations.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"214 ","pages":"Article 106640"},"PeriodicalIF":4.0,"publicationDate":"2025-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144865588","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}