Insect Biochemistry and Molecular Biology最新文献

{"title":"Structural and mechanistic insights into BoCSP1-mediated resistance to neonicotinoids in Bradysia odoriphaga","authors":"Xingyu Ma,&nbsp;Junjie Zeng,&nbsp;Chunni Zhang,&nbsp;Wu Dai","doi":"10.1016/j.ibmb.2026.104489","DOIUrl":"10.1016/j.ibmb.2026.104489","url":null,"abstract":"<div><div>The chemosensory protein (CSP)-mediated sequestration resistance mechanism has attracted increasing attention. This study revealed that a laboratory-selected clothianidin-resistant strain of <em>Bradysia odoriphaga</em> exhibited moderate cross-resistance to thiamethoxam. qPCR analysis demonstrated significant upregulation of <em>BoCSP1</em> expression in the resistant strain, and its transcription could be induced by clothianidin exposure. RNAi-mediated silencing of <em>BoCSP1</em> increased the susceptibility of resistant larvae to both clothianidin and thiamethoxam. Fluorescence competitive binding assays showed strong binding affinity between BoCSP1 and clothianidin/thiamethoxam, with <em>Ki</em> values &lt; 3 μM. Molecular dynamics simulations and binding mode analysis further revealed that the stability of BoCSP1-clothianidin and BoCSP1-thiamethoxam complexes is primarily governed by van der Waals interactions. Computational alanine scanning (CAS) and site-directed mutagenesis identified Asn74 and Ile90 as critical residues mediating BoCSP1-clothianidin binding, while Trp101 was essential for BoCSP1-thiamethoxam interaction. Binding pocket analysis of the mutant proteins revealed that alanine substitutions disrupted hydrogen bonding networks and altered local conformational rigidity, leading to substantially reduced ligand affinity. These findings elucidate a novel CSPs-mediated resistance mechanism and provide a theoretical foundation for developing precision-engineered insecticides that exploit CSPs as unique molecular targets.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"188 ","pages":"Article 104489"},"PeriodicalIF":3.7,"publicationDate":"2026-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145916318","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A cuticular protein essential for sperm dimorphism and male fertility in Bombyx mori","authors":"Zhongwei Li ,&nbsp;Bing Zhang ,&nbsp;Chenyue Zhao ,&nbsp;Yanyan Wang ,&nbsp;Ruoke Wang ,&nbsp;Camilo Ayra-Pardo ,&nbsp;Yunchao Kan ,&nbsp;Dandan Li","doi":"10.1016/j.ibmb.2025.104476","DOIUrl":"10.1016/j.ibmb.2025.104476","url":null,"abstract":"<div><div>In Lepidoptera, sperm develop as two distinct types: fertile eupyrene and non-fertile apyrene sperm. Although apyrene sperm do not fertilize eggs, they support fertilization through auxiliary functions. However, the molecular mechanisms underlying this dimorphism remain poorly understood. Here, we identify the Cuticular Protein Hypothetical 19 precursor (<em>BmorCPH19</em>) gene as essential for sperm development in <em>Bombyx mori</em>. Using 70 μm cell strainers, we isolated transcriptionally active apyrene spermatocytes and an eupyrene-enriched fraction containing mature sperm bundles from 3-day-old pupal testes, when morphs are distinct and separable. Transcriptomic profiling revealed 737 differentially expressed genes, including <em>BmorCPH19</em>, unexpectedly enriched in the eupyrene fraction —suggesting a novel role beyond cuticle-related activity. qRT-PCR analysis showed that <em>BmorCPH19</em> expression in the testes peaks during eupyrene differentiation in the 6-day-old fourth-instar larva and the 1-day-old prepupa. Isolated sperm morphs confirmed consistent <em>BmorCPH19</em> enrichment in eupyrene sperm, with maximal accumulation in late pupae. Functional assays demonstrated its role in sperm fate: RNAi knockdown increased the apyrene-to-eupyrene ratio, while overexpression favored eupyrene sperm but impaired fertility, yielding more non-viable eggs. These findings identify BmorCPH19 as the first cuticular protein essential for lepidopteran sperm dimorphism and reveal its dosage-sensitive role in balancing sperm differentiation and reproductive success, offering a potential molecular target for pest control.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"188 ","pages":"Article 104476"},"PeriodicalIF":3.7,"publicationDate":"2025-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145883281","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Two cytochrome P450 epoxidases mediate juvenile hormone biosynthesis in Drosophila melanogaster","authors":"Daiki Fujinaga ,&nbsp;Yuya Ohhara ,&nbsp;Naoki Okamoto ,&nbsp;Hannah Chu ,&nbsp;Kerry E. Mauck ,&nbsp;Naoki Yamanaka","doi":"10.1016/j.ibmb.2025.104475","DOIUrl":"10.1016/j.ibmb.2025.104475","url":null,"abstract":"<div><div>Juvenile hormones (JHs) mediate various biological processes such as development and reproduction in insects. Although pleiotropic functions of JHs are well investigated in the fruit fly <em>Drosophila melanogaster</em>, their biosynthetic mechanisms are less well understood, partly because many JH biosynthetic enzymes still remain unidentified in this important model species. Here we report that two cytochrome P450 (CYP) epoxidases mediate JH biosynthesis in <em>D. melanogaster</em>. In addition to previously reported Cyp6g2, a second epoxidase, Cyp6a13, also functions in the corpus allatum, the major JH biosynthetic endocrine gland. Combined mutations of the genes encoding these enzymes cause developmental and reproductive defects, which can be rescued by JH application. JH biosynthetic functions of these genes were further confirmed by using a heterologous expression system and <em>ex vivo</em> tissue culture. Collectively, our results indicate that these two CYP epoxidases function cooperatively to mediate JH biosynthesis in <em>D. melanogaster</em>.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"187 ","pages":"Article 104475"},"PeriodicalIF":3.7,"publicationDate":"2025-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145880990","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Novel-miR-932 targets BgNeverland to regulate 20-hydroxyecdysone synthesis in Blattella germanica: Prospective target for pest control","authors":"Yue Zhang ,&nbsp;Fei Yu ,&nbsp;Xian Huang ,&nbsp;Xiaoying Lv ,&nbsp;Xunxun Wang ,&nbsp;Zexin Gao ,&nbsp;Xianyi Wang","doi":"10.1016/j.ibmb.2025.104474","DOIUrl":"10.1016/j.ibmb.2025.104474","url":null,"abstract":"<div><div>miRNAs critically regulate hormonal signaling to ensure precise spatiotemporal control of metamorphosis and reproduction in insects, highlighting their potential as pest control targets. <em>Blattella germanica</em>, is a major indoor sanitary pest that threatens human and domestic animal health. However, the miRNA expression profiles and their systematic functions in the hormonal signaling pathways remain largely uncharacterized in <em>B. germanica</em>. Transcriptomic analysis of the nymph-to-adult transition in <em>B. germanica</em> revealed that both downregulated differentially expressed genes and the target genes of differentially expressed miRNAs were significantly enriched in insect hormone biosynthesis pathways. RNA interference (RNAi)-mediated silencing of six key 20E biosynthesis genes (<em>BgNeverland</em>, <em>BgSpook</em>, <em>BgPhantom</em>, <em>BgDisembodied</em>, <em>BgShadow</em>, and <em>BgShade</em>) resulted in molting defects and increased mortality in nymphs. Based on bioinformatic predictions and omics data, we identified miRNAs potentially targeting 20E biosynthesis genes. Subsequent dual-luciferase reporter assays, along with agomir and antagomir injection experiments, confirmed that novel-miR-932 negatively regulates <em>BgNeverland</em>. Both oversupply and inhibition of novel-miR-932 led to elevated nymphal mortality. Supplementation with exogenous 20E effectively rescued the increased mortality induced by novel-miR-932 agomir injection. Furthermore, either oversupply of miR-932 or silencing of <em>BgNeverland</em> resulted in a reduction in lipid content, consequently compromising female fecundity. These results implicate miRNAs that target 20E pathway genes as potential targets for novel pest control strategies.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"188 ","pages":"Article 104474"},"PeriodicalIF":3.7,"publicationDate":"2025-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145861668","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Odorant receptor CaspOR39 underpinned cineole-mediated oviposition repellency in the rape stem weevil Ceutorhynchus asper","authors":"Fuqiang Rao ,&nbsp;Jie Yang ,&nbsp;Danhao Peng ,&nbsp;Xinghao Li ,&nbsp;Zhaoqun Li ,&nbsp;Deguang Liu","doi":"10.1016/j.ibmb.2025.104473","DOIUrl":"10.1016/j.ibmb.2025.104473","url":null,"abstract":"<div><div>Herbivorous insects rely on olfactory cues to select oviposition sites on host plants, which is a process critical for their reproductive success. The rape stem weevil, <em>Ceutorhynchus asper</em> Roel. (Coleoptera: Curculionidae), is a major pest that causes significant damage to rapeseed and other cruciferous crop production, yet the chemical and molecular mechanisms underlying its oviposition preference remain unknown. Our behavioral assays revealed that <em>C. asper</em> females strongly avoided pre-infested rapeseed plants. Five monoterpenes (p-cymene, 1,8-cineole, ocimene, camphor, and α-pinene) were identified as herbivore-induced plant volatiles of <em>C. asper</em>-infested plants, with their emission levels significantly higher than those in healthy plants. Among these five compounds, subsequent electroantennogram (EAG) screening revealed that only 1,8-cineole triggered significant responses in <em>C. asper</em> and showed a concentration-dependent sensitivity. Behavioral assays showed that female <em>C. asper</em> exhibited significant behavioral and oviposition avoidance when exposed to 1,8-cineole. We identified CaspOR39 as the specific odorant receptor for 1,8-cineole, with molecular docking for prediction of the binding affinity and key residues. RNAi of <em>CaspOR39</em> abolished female EAG responses, as well as behavioral and oviposition avoidance of 1,8-cineole. These results revealed a sex-specific chemical sensing mechanism: female <em>C. asper</em> can utilize CaspOR39 to detect 1,8-cineole as a key cue of plant infestation, driving adaptive oviposition behaviors of this beetle. Our results provide mechanistic and molecular insights into CaspOR39-mediated oviposition behaviors of <em>C. asper</em>, and a basis for the development of semiochemical-based strategies for management of this weevil.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"187 ","pages":"Article 104473"},"PeriodicalIF":3.7,"publicationDate":"2025-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145861595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CRISPR/Cas9-based evidence that overexpression of Gm-mGST1 mediates abamectin resistance in the oriental fruit moth, Grapholita molesta","authors":"Sha Su ,&nbsp;Xiaohe Zhang ,&nbsp;Xinyu Wang ,&nbsp;Caiyun Qiu ,&nbsp;Zhimin Xu ,&nbsp;Jaime C. Piñero ,&nbsp;Xiong Peng ,&nbsp;Fei Li ,&nbsp;Yayun Zuo ,&nbsp;Maohua Chen","doi":"10.1016/j.ibmb.2025.104472","DOIUrl":"10.1016/j.ibmb.2025.104472","url":null,"abstract":"<div><div>Abamectin-based insecticides are widely used in integrated pest management and are particularly effective against fruit borers such as the oriental fruit moth, <em>Grapholita molesta</em>. However, rapid resistance evolution threatens their long-term efficacy. This study elucidates the role of the glutathione S-transferase gene <em>Gm-mGST1</em> in abamectin resistance in <em>G. molesta</em>. A laboratory-selected resistant strain (AB-R) exhibited an 85.5-fold increase in resistance compared with a susceptible strain (AB-S). Sequencing of glutamate-gated chloride channel (<em>GmGluCl</em>) gene revealed no target-site mutations, implicating a metabolic resistance mechanism. In AB-R, GST enzymatic activity was significantly elevated. GST synergist diethyl maleate (DEM) increased the toxicity of abamectin more strongly in the abamectin-resistant <em>G. molesta</em> strain than in the susceptible strain, indicating that GSTs contribute to abamectin resistance. <em>Gm-mGST1</em> showed strong and stage-specific overexpression under abamectin exposure. Functional analysis using CRISPR/Cas9 knockout of <em>Gm-mGST1</em> in the AB-R strain reduced resistance 16.3-fold, providing the definitive evidence that a <em>GST</em> gene directly mediates abamectin resistance in <em>G. molesta</em>. The catalytic activity of recombinant Gm-mGST1 was verified <em>in vitro</em> using CDNB as the substrate. Additionally, abamectin exhibited a certain degree of inhibitory effect on the activity of Gm-mGST1. HPLC analysis further revealed that the peak area of abamectin significantly decreased in the presence of recombinant Gm-mGST1, while ectopic expression in <em>Drosophila melanogaster</em> increased abamectin tolerance by 1.97-fold. There is a significant positive correlation between the abamectin resistance levels and the expression levels of <em>Gm-mGST1</em> in field populations of <em>G. molesta</em>. These findings identify <em>Gm-mGST1</em> as a critical gene involved in abamectin resistance and establish it as a potential molecular marker for monitoring resistance in field populations. More broadly, this study sets a precedent for integrating CRISPR/Cas9 gene editing into insecticide resistance research, bridging the gap between correlative evidence and functional validation, and providing a framework for developing GST-targeted resistance management strategies in orchard pests. This study provides evidence using CRISPR/Cas9 to confirm the contribution of GST to abamectin resistance in insects.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"187 ","pages":"Article 104472"},"PeriodicalIF":3.7,"publicationDate":"2025-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145852868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcription factor Schlank positively regulates odorant-binding protein 9 expression to mediate recognition of two volatiles in Nilaparvata lugens (Stål)","authors":"Rui-Lin Zhang,&nbsp;Hao-Wei Wang,&nbsp;Man-Qun Wang","doi":"10.1016/j.ibmb.2025.104471","DOIUrl":"10.1016/j.ibmb.2025.104471","url":null,"abstract":"<div><div>Odorant-binding proteins (OBPs) are crucial components of the insect olfactory system, serving as the initial step in peripheral olfactory signal transduction. They mediate odorant detection by selectively binding and transporting volatile chemical cues through the sensillum lymph to olfactory receptors. To elucidate the transcriptional mechanisms that regulate insect olfactory gene expression, this study focuses on the <em>NlOBP9</em> gene in the brown planthopper, <em>Nilaparvata lugens</em>. Here, we employed dual-luciferase reporter assays and yeast one-hybrid (Y1H) experiments to demonstrate that the transcription factor Schlank specifically binds to the promoter region of <em>NlOBP9</em>, thereby modulating its transcriptional activity. Tissue-specific expression profiling revealed that Schlank is highly expressed in the antennae of female adults. RNA interference (RNAi)-mediated silencing of <em>Schlank</em> significantly reduced <em>NlOBP9</em> expression and abolished the avoidance behaviour of female adults towards linalool and methyl benzoate. This study provides the first evidence that Schlank specifically regulates members of the <em>NlOBPs</em> family within the olfactory system, offering new molecular insights into the precise regulatory mechanisms underlying insect olfactory gene expression.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"187 ","pages":"Article 104471"},"PeriodicalIF":3.7,"publicationDate":"2025-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145814929","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sgie3, a venomous carboxylesterase from the ectoparasitoid of Sclerodermus guani exhibits lipolytic activity to modulate host lipid metabolism","authors":"Chaoyan Wu ,&nbsp;Yuhan Liu ,&nbsp;Jianfei Dao ,&nbsp;Tao Zhu ,&nbsp;Guoxing Wu ,&nbsp;Jiaying Zhu","doi":"10.1016/j.ibmb.2025.104470","DOIUrl":"10.1016/j.ibmb.2025.104470","url":null,"abstract":"<div><div>Carboxylesterases are commonly found in the venom of parasitoids, yet their physiological roles as venom proteins in parasitoids remain to be elucidated. This study identified 19 carboxylesterase genes within the genome of <em>Sclerodermus guani</em>, with Sgie3 showing specific and abundant expression in the venom gland, implicating it as a key venom constituent with a vital role in host regulation. Functional characterization demonstrated that Sgie3 displays hydrolytic activity towards triglycerides <em>in vitro</em>. Injection of Sgie3 induces triglyceride increase and reduction in lipid droplet size in <em>Tenebrio molitor</em> pupae, the host of <em>S. guani</em>, resulting in dose-dependent developmental arrest effects in the pupae. Lipidomic analysis indicated that Sgie3 injection altered 436 lipid metabolites after one day, notably enriched in glycerophospholipid, arachidonic acid, α-linolenic acid, glycerolipid metabolism, as well as fatty acid elongation and degradation, and fatty acid autophagy pathways. These findings highlight the specific role of Sgie3 in modulating host lipid metabolism, providing insights into how parasitoids employ venom to manipulate host nutritional metabolism to support the successful development of their offsprings.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"187 ","pages":"Article 104470"},"PeriodicalIF":3.7,"publicationDate":"2025-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145792825","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Antiviral RNAi is preserved in dsRNA-resistant Colorado potato beetle (Leptinotarsa decemlineata)","authors":"Luis Hernández-Pelegrín ,&nbsp;Swati Mishra ,&nbsp;Juan Luis Jurat-Fuentes ,&nbsp;Salvador Herrero","doi":"10.1016/j.ibmb.2025.104469","DOIUrl":"10.1016/j.ibmb.2025.104469","url":null,"abstract":"<div><div>Biocontrol strategies based on RNA interference (RNAi) hold great potential against the Colorado potato beetle (<em>Leptinotarsa decemlineata</em>), but the emergence of dsRNA-resistant populations threatens their sustained efficacy. In insects, RNAi constitutes a major antiviral defense mechanism, primarily through the production of virus-derived small interfering RNAs (vsiRNAs) and, in certain species, PIWI-interacting RNAs (vpiRNAs). Certain viruses can muffle this response by expressing viral suppressors of RNAi (VSRs). We hypothesized that viruses encoding VSRs may contribute to the high resistance levels to dsRNA reported in a population of <em>L. decemlineata</em> (CEAS 300). In testing this hypothesis, we first characterized the virome of two susceptible and the dsRNA-resistant CEAS 300 <em>L. decemlineata</em> populations and identified eight novel RNA viruses. Comparative analysis revealed no association between RNA virus abundance and incidence among the populations and the dsRNA resistance phenotype. Small RNA profiling revealed a combination of vsiRNAs (widespread) and vpiRNAs (against a single virus) in all three <em>L. decemlineata</em> populations, confirming that the antiviral RNAi machinery remains active in the dsRNA-resistant population and that resistance is not due to defects in core RNAi components. Conversely, dsRNA treatment affected the abundance and vsiRNA response to one virus, and these effects were predominantly observed in the dsRNA-resistant population. Overall, our findings provide new insights into resistance to RNAi and host-virus interactions, highlighting the relevance of RNA viruses in the context of RNAi-based pest control strategies.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"187 ","pages":"Article 104469"},"PeriodicalIF":3.7,"publicationDate":"2025-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145773199","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expanded UDP-glycosyltransferase gene clusters underlie insecticide detoxification in Spodoptera frugiperda","authors":"Yajie Kong ,&nbsp;Xing Geng ,&nbsp;Huiru Zhou ,&nbsp;Xinyue Su ,&nbsp;Kairan Zuo ,&nbsp;Huidong Wang ,&nbsp;Angela Hayward ,&nbsp;Chris Bass ,&nbsp;Shutang Zhou","doi":"10.1016/j.ibmb.2025.104468","DOIUrl":"10.1016/j.ibmb.2025.104468","url":null,"abstract":"<div><div>As Phase II detoxification enzymes, UDP-glycosyltransferases (UGTs) catalyze the conjugation of sugar moieties to various hydrophobic xenobiotics, thereby increasing their solubility and facilitating excretion. Certain UGT gene families are often massively expanded as clusters in insect genomes, yet whether such expansions confer a preadaptive advantage for insecticide detoxification remains unclear. In this study, we employed three previously generated <em>Spodoptera frugiperda</em> knockout strains, in which three UGT gene clusters—comprising ten and four UGT33 genes and fifteen UGT40 genes—were deleted using CRISPR-Cas9. By comparing the insecticide susceptibility of these knockout strains to that of the background strain across six insecticides with distinct modes of action, we found that the UGT40 gene cluster contributes to tolerance to emamectin benzoate and lambda-cyhalothrin. Further functional assays using transgenic <em>Drosophila melanogaster</em> lines overexpressing each of the 15 UGT40 genes identified five genes involved in insecticide detoxification, including four associated with emamectin benzoate (<em>UGT40R14</em>, <em>UGT40F19</em>, <em>UGT40M10</em>, and <em>UGT40L10</em>) and one (<em>UGT40R17</em>) with lambda-cyhalothrin. Computational modelling of protein structure and ligand binding analyses suggested that, in the case of the UGTs associated with emamectin benzoate tolerance, the main component of the insecticide, emamectin B1a, binds in a position where the key interactions are with the N-terminal domain, an orientation with the sugar acceptor and metabolic activity. Together, our results provide a systematic dissection of UGT gene cluster function in insecticide detoxification and highlight UGTs as an underappreciated but important component of metabolic resistance in agricultural pests.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"187 ","pages":"Article 104468"},"PeriodicalIF":3.7,"publicationDate":"2025-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145720175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}