Insect Biochemistry and Molecular Biology最新文献

{"title":"Drosophila proteasome subunit Rpn8 controls IMD pathway activation via PGRP-SC2 degradation","authors":"Mingfei Wu ,&nbsp;Umar Ali ,&nbsp;Sen Ding ,&nbsp;Chuchu Zhang ,&nbsp;Yiheng Jin ,&nbsp;Xinyan Cao ,&nbsp;Muhammad Abdul Rehman Saeed ,&nbsp;Yangyang Zhu ,&nbsp;Muhammad Usama ,&nbsp;Badar Raza ,&nbsp;Qingshuang Cai ,&nbsp;Shanming Ji","doi":"10.1016/j.ibmb.2025.104422","DOIUrl":"10.1016/j.ibmb.2025.104422","url":null,"abstract":"<div><div>The ubiquitin-proteasome system (UPS) is a highly conserved protein degradation pathway in insects, playing crucial roles in cellular homeostasis, signal transduction, and various biological processes. Here, we demonstrate that the <em>Drosophila melanogaster</em> (fruit fly) regulatory particle non-ATPase 8 (Rpn8), a key subunit of the 26 S proteasomal cap, is indispensable for fly survival upon bacterial infections. Through targeted genetic knockdown, we observed that <em>Rpn8</em>-silenced flies exhibited decreased expressions of several antimicrobial peptides upon bacterial challenges, concomitant with impaired bacterial clearances. Further proteomic and biochemical analyses revealed that Rpn8 controls the proteasome-mediated degradation of peptidoglycan recognition protein SC2 (PGRP-SC2), a typical amidase that antagonizes immune deficiency (IMD) signaling. Silencing of <em>PGRP-SC2</em> prevented the functional role of <em>Rpn8</em> in controlling the fly antibacterial immune defense, suggesting that <em>Rpn8</em> mediates <em>Drosophila</em> innate immunity in a <em>PGRP-SC2</em>-dependent manner. Intriguingly, overexpression of <em>Bombyx mori</em> (silkworm) <em>Rpn8</em> rescued the immune defects in <em>Rpn8 RNAi</em> flies after bacterial infections. Collectively, our findings not only establish <em>Rpn8</em> as a novel immune modulator that bridges UPS-mediated protein degradation with innate immune defense in <em>Drosophila</em> but also offer insights into conserved mechanisms of host-pathogen interactions in other insect species.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"185 ","pages":"Article 104422"},"PeriodicalIF":3.7,"publicationDate":"2025-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145285149","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 conserved Piezo–mRPL2/Jun axis controls reproduction via mitochondrial bioenergetics","authors":"Jiangwen Luo ,&nbsp;Yaru Lu ,&nbsp;Zhiruo Zhang ,&nbsp;Qian Zhang,&nbsp;Yunzhu Shang,&nbsp;Yajie Yuan,&nbsp;Xiaohui Duan,&nbsp;Long Zhang,&nbsp;Yuxia Tang,&nbsp;Hai Hu,&nbsp;Xiaoling Tong,&nbsp;Fangyin Dai","doi":"10.1016/j.ibmb.2025.104417","DOIUrl":"10.1016/j.ibmb.2025.104417","url":null,"abstract":"<div><div>Piezo channels are essential mechanosensors, but their roles in reproduction remain poorly understood. Here, we identify a critical function for BmPiezo in silkworm fertility and embryonic diapause, mediated through a conserved downstream regulatory axis. CRISPR/Cas9-mediated knockout of BmPiezo resulted in male sterility and loss of embryonic diapause, accompanied by dysregulation of ribosome-associated genes and oxidative phosphorylation, and reduced ATP production. We identified mitochondrial ribosomal protein L2 (BmmRPL2) and transcription factor BmJun as key downstream effectors, required for spermatogenesis and diapause, respectively. Homozygous disruption of either gene caused embryonic lethality. Cross-species expression analysis revealed strong expression of Piezo2 and mRPL2 in early human spermatids. Remarkably, transgenic expression of murine Piezo2 in BmPiezo−/−silkworms reactivated the Piezo–mRPL2/Jun axis, restored ATP levels, and rescued reproductive defects. These findings define a conserved pathway that connects mechanical sensing to mitochondrial bioenergetics in reproductive regulation, advancing our understanding of fertility mechanisms across species.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"185 ","pages":"Article 104417"},"PeriodicalIF":3.7,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145278602","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":"Bombyx mori nucleopolyhedrosis virus-derived circular RNAs with protein-coding potential facilitate viral replication","authors":"Xing Zhang ,&nbsp;Zeen Shen ,&nbsp;Yani Kang ,&nbsp;Wenbin Yu ,&nbsp;Xiaoyan Du ,&nbsp;Qian Teng ,&nbsp;Zihan He ,&nbsp;Chengliang Gong ,&nbsp;Xiaolong Hu","doi":"10.1016/j.ibmb.2025.104420","DOIUrl":"10.1016/j.ibmb.2025.104420","url":null,"abstract":"<div><div>Circular RNAs (circRNAs) are renowned for their exceptional stability and have been increasingly recognized for their dual roles in pro- and antiviral responses during host-virus interactions. In this study, we investigated the functional landscape of circular RNAs (circRNAs) during infection with <em>Bombyx mori</em> nucleopolyhedrosis virus (BmNPV), a model baculovirus system. Our comprehensive analysis revealed that hundreds of host-derived circRNAs are dynamically regulated upon BmNPV infection, while the virus itself generates numerous viral circRNAs (vcircRNAs) via back-splicing. Using a combination of advanced experimental approaches, we validated the existence of multiple cellular and viral circRNAs. Among these, we characterized vcircRNA-390, a BmNPV-encoded circRNA harboring a small open reading frame (ORF) and four viral internal ribosome entry sites (IRESs). Remarkably, vcircRNA-390 serves as a template for the translation of an 81-amino acid viral peptide (VSP81). Functional studies demonstrated that both vcircRNA-390 and VSP81 significantly enhance viral replication. Mechanistically, we provide evidence that VSP81 likely targets the host RNA interference (RNAi) pathway, a major antiviral defence system, to promote viral immune evasion. While these findings establish vcircRNA-390 as a novel proviral factor in insect-virus interactions, the detailed molecular mechanisms by which VSP81 modulates the RNAi machinery remain to be fully elucidated. Our work not only expands the understanding of viral circRNA biology but also reveals a new dimension of the host-pathogen conflict, wherein BmNPV exploits circRNA-mediated translation to subvert antiviral defences.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"185 ","pages":"Article 104420"},"PeriodicalIF":3.7,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145273296","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":"Assessment of RNA interference effectiveness mediated by siRNA sequences and structures in Drosophila S2 cells","authors":"Hyun-Soo Kim ,&nbsp;Huipeng Pan ,&nbsp;Subba Reddy Palli ,&nbsp;June-Sun Yoon","doi":"10.1016/j.ibmb.2025.104418","DOIUrl":"10.1016/j.ibmb.2025.104418","url":null,"abstract":"<div><div>Extensive research has clarified the mechanisms of RNA interference (RNAi) in insects. Although double-stranded RNA (dsRNA) effectively induces RNAi in insects, comprehensive studies on how small interfering RNA (siRNA) structural characteristics influence messenger RNA (mRNA) cleavage efficiency are limited. This study systematically examined the impact of diverse sequence and structural modifications on the efficiency of siRNA-mediated knockdown of target genes in <em>Drosophila melanogaster</em>. Our findings indicate that siRNA efficacy drastically decreased at a length of 17 nucleotides (nt), but was restored by extending to 19 base pairs (bp) with random sequences. Additionally, siRNAs with 2-nt overhangs demonstrated greater efficacy compared to blunt-ended structures. We found that the knockdown efficiency varies depending on the secondary structure characteristics of the mRNA region to which siRNA binds. Furthermore, next-generation sequencing was employed to map predicted siRNA distributions to actual dsRNA processing patterns, allowing detailed profiling of cleavage depth and sequence preferences. Collectively, these results enhance our understanding of the relationship between siRNA sequence design parameters and RNAi efficiency in <em>Drosophila</em>, providing a significant advance in siRNA-based RNAi research in insects. This study also offers valuable insights into dsRNA off-target effects.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"185 ","pages":"Article 104418"},"PeriodicalIF":3.7,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145256973","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":"Apis mellifera multi-omics responses to ectoparasitic mites: unveiling biomarkers for Tropilaelaps mercedesae and Varroa destructor in honeybee hemolymph through hemolymph peptide and proteome analysis using MALDI-MS and LC-ESI-MS/MS","authors":"Patcharin Phokasem ,&nbsp;Michel Bocquet ,&nbsp;Sébastien N. Voisin ,&nbsp;Chainarong Sinpoo ,&nbsp;Camille Houdelet ,&nbsp;Panuwan Chantawannakul ,&nbsp;Terd Disayathanoowat ,&nbsp;Philippe Bulet","doi":"10.1016/j.ibmb.2025.104416","DOIUrl":"10.1016/j.ibmb.2025.104416","url":null,"abstract":"<div><div><em>Varroa destructor</em> and <em>Tropilaelaps mercedesae</em> are ectoparasites that pose a significant threat to honeybee populations. It is essential to understand their impact to develop effective management strategies. In this study, we examined the relative abundance of proteins and the expression of immune-related genes in <em>A. mellifera</em> parasitized by the two mites using mass spectrometry analysis of hemolymph and transcriptomics of bleeding honeybees. The study revealed the modulation of genes and proteins associated with immune response and stress management. The <em>apidaecin</em> gene was upregulated in <em>Varroa</em>-infested honeybees and downregulated in <em>Tropilaelaps</em>-infested honeybees, a finding that correlated with MALDI-MS data. Furthermore, variations in the abundance of Hexamerin, Odorant Binding Protein 21, Phenoloxidase and Transferrin were observed in <em>Varroa-</em> and <em>Tropilaelaps</em>-infested honeybees. Using proteomics, potential markers associated with immunity, metabolism and stress responses (e.g. high Glx storage protein, alpha-glucosidase, glyoxylate/hydroxypyruvate reductases) were found to be specific to <em>Varroa</em> and <em>Tropilaelaps</em> infestations. Two Deformed Wing Virus (DWV) proteins exhibit higher abundance in the hemolymph of <em>Tropilaelaps-</em>infested honeybees compared to non-infested and <em>Varroa</em>-infested honeybees, suggesting that <em>Tropilaelaps</em> may weaken honeybee immunity, potentially exacerbating viral infections and colony health decline. This indicates that studying host-mite species interactions could lead to a better understanding of honeybee protection.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"185 ","pages":"Article 104416"},"PeriodicalIF":3.7,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145256909","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":"Kinetic studies of pheromone-binding proteins (LdisPBP1 and LdisPBP2) from spongy moth Lymantria dispar with natural ligands and their analogues: Enantiomer distinction at external and internal binding sites","authors":"Erika Plettner,&nbsp;Ashna Aulakh ,&nbsp;Mailyn Terrado","doi":"10.1016/j.ibmb.2025.104419","DOIUrl":"10.1016/j.ibmb.2025.104419","url":null,"abstract":"<div><div>Olfaction plays an important role in the reproduction of the spongy moth, <em>Lymantria dispar</em>, a major defoliator of deciduous trees. Detection of the attractant sex pheromone, (7<em>R</em>, 8<em>S</em>)-epoxy-2-methyloctadecane, (+)-disparlure, aids males in the location of mates by triggering upwind flight behavior. The antipode, (−)-disparlure, inhibits upwind flight behavior. The sensory hairs on the antennae of the male moth are innervated by the dendrite of an olfactory neuron, which is immersed in sensory lymph, a solution with pheromone-binding proteins (PBPs). These proteins bind pheromone molecules selectively and are necessary for proper olfactory functioning. Two PBPs from spongy moth are known: <em>LdisPBP1</em> and <em>LdisPBP2</em>. The two proteins exhibit differences in their binding affinities towards various natural and unnatural ligands at their internal binding sites. Ligands are bound by PBPs in a two-step process: ligand capture results in binding to an external site, which is followed by movement of the ligand to the internal site. In this study, we investigate the differences in association and dissociation rates of the two proteins with (+)-disparlure, its enantiomer, (−)-disparlure, and some analogues with heteroatom substitutions of oxygen or sulfur at either carbon-5 or carbon-10, using stopped-flow kinetics. We used a fluorescent reporter (N-phenyl-1-naphthylamine, NPN), which competes with the ligands of interest and reports on external site binding in PBPs. The results are discussed in the context of kinetic selectivity of PBPs towards natural disparlure and synthetic analogue ligands, at their external binding sites, where enantiomer distinction has not been detected previously. Ligand docking simulations at two external sites of A and B forms and at one internal site of the B forms of the two proteins were used to interpret the enantioselectivity in the association and dissociation of the ligands. This paper also discusses the enantiomer selectivity and the molecular role of PBPs in the desorption from hydrophobic surfaces and partition between lipid vesicles and aqueous solutions of the pheromone enantiomers and their heteroatom-substituted analogues. Results are interpreted in terms of the contribution of ligand association and dissociation kinetics to the enantiomer selectivity of the spongy moth PBPs.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"185 ","pages":"Article 104419"},"PeriodicalIF":3.7,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145273206","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":"Female accessory reproductive glands of Paederus fuscipes serve as a reservoir of symbiotic pederin-producing bacteria","authors":"Xuhao Song ,&nbsp;Hui Meng ,&nbsp;Tingbang Yang ,&nbsp;Yujie Li ,&nbsp;Fake Zheng ,&nbsp;Xianghui Yan","doi":"10.1016/j.ibmb.2025.104408","DOIUrl":"10.1016/j.ibmb.2025.104408","url":null,"abstract":"<div><div><em>Paederus fuscipes</em>, an ecologically and medically important species, is known for its blistering toxin pederin in hemolymph. Evidence demonstrates that the toxin is synthesized by the uncultured symbiotic pederin-producing bacteria (PPB) in <em>P. fuscipes</em>, but the biological characteristics of PPB within the beetle host remain poorly characterized. Here, we investigated PPB abundance variations in <em>P. fuscipes</em> across different factors (sexes, life stages, habitats, and organs), along with their colonization sites and metabolic potentials. The findings revealed that the PPB abundance in female <em>P. fuscipes</em> at the level of individuals and tissues exhibited stable colonization patterns, independent of habitat and time changes. Notably, PPB dominated the bacterial community in females (relative abundance ≥66.08 %) and nearly occupied reproductive organs (relative abundance ≥96.31 %). Moreover, our results indicated that PPB were predominantly enriched in the accessory glands of female reproductive organs, which could serve as a reservoir for PPB proliferation. Although PPB were not cultured in this study, metagenomic binning yielded the draft genome of PPB (CheckM completeness = 85.14 %, contamination = 0), in which genes related to pederin biosynthesis were identified. Phylogenetic analyses revealed that PPB formed a sister clade to <em>Pseudomonas aeruginosa</em> rather than nesting within the <em>P. aeruginosa</em> lineage. Metabolic module prediction analysis revealed specific deficiencies in PPB's energy metabolism and amino acid biosynthesis pathways, suggesting limited free-living potential for PPB. Collectively, this study provides insights into PPB biological characteristics within their beetle host and paves the way for biotechnological exploitation related to pederin production.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"185 ","pages":"Article 104408"},"PeriodicalIF":3.7,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145228382","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":"Determining the type of aerobic/anaerobic metabolism during exercise in Drosophila using incremental loading exercise","authors":"Qin Yi ,&nbsp;Meng Ding ,&nbsp;Jinglin Liu,&nbsp;Xiaoya Wang,&nbsp;Lan Zheng","doi":"10.1016/j.ibmb.2025.104404","DOIUrl":"10.1016/j.ibmb.2025.104404","url":null,"abstract":"<div><div>Currently, <em>Drosophila</em> exercise is only classified as regular or endurance exercise, and there is no aerobic or anaerobic form of exercise, making <em>Drosophila</em> exercise modeling studies limited. Therefore, this study aimed to determine the type of aerobic and anaerobic exercise in <em>Drosophila</em> using Incremental Load exercise. A seven-day-old wild-type <em>Drosophila</em> was used as a study sample for exercise intervention using a <em>Drosophila</em> exercise device.</div><div>This study was designed with six exercise intensities, including 0.31 rev/s (E1), 0.45 rev/s (E2), 0.59 rev/s (E3), 0.71 rev/s (E4), 0.83 rev/s (E5), 0.91 rev/s (E6), and Control in addition <em>Drosophila</em> in all exercise groups performed a one-time 2.5-h acute exercise, and the control group did no exercise intervention. Anaerobic and aerobic metabolic enzymes in <em>Drosophila</em> were assayed immediately after exercise. Additionally, <em>Drosophila</em> exercise types were determined using hemolymph lactate, respiratory quotient, trehalose concentration, and mitochondrial respiration assays.</div><div>The experimental results show that at E2 exercise intensity, <em>Drosophila</em> mainly uses aerobic metabolism, and at E6 exercise intensity, it mainly uses anaerobic metabolism.We exercised <em>Drosophila</em> for 2 weeks at E2 and E6 exercise intensities to examine whether the physiological differences (cardiac function, climbing ability, sleep ability, lifespan, etc.) were consistent with those observed in mammals undergoing long-term aerobic and anaerobic exercise.The results showed that these differences are consistent with mammals.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"185 ","pages":"Article 104404"},"PeriodicalIF":3.7,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145224667","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":"The role of the transformer gene in sex determination and its employment in CRISPR/Cas9-based homing gene drive in the global fruit pest Drosophila suzukii","authors":"Dan Deng ,&nbsp;Xueying Yi ,&nbsp;Wen Wen,&nbsp;Liuqing He,&nbsp;Wei Peng","doi":"10.1016/j.ibmb.2025.104406","DOIUrl":"10.1016/j.ibmb.2025.104406","url":null,"abstract":"<div><div>Sex determination of Diptera is established by the cascade genes such as <em>transformer</em> (<em>tra</em>), though the primary signals for sex determination differ among different insects. Here, we report the isolation, expression and function of <em>tra</em> gene in an invasive pest, <em>Drosophila suzukii</em>, and study the potential use of the <em>D. suzukii tra</em> (<em>Dstra</em>) gene in CRISPR/Cas9-based homing gene drive for genetic-based pest management. The <em>Dstra</em> gene is highly conserved in structure and has a sex-specific transcript. To test the function of this gene in sex determination, <em>Dstra</em> dsRNA was injected into embryos. Almost all XX embryos developed into masculinized phenotypic male adults with intersex morphology. Abnormal ovaries were revealed in XX pseudomales upon dissection. Based on the necessary role of <em>Dstra</em> for female development, we developed and evaluated a homing gene drive that targets <em>Dstra</em> in <em>D</em>. <em>suzukii</em>. The drive component consisting of multiplex <em>Dstra</em> single guide RNAs and Cas9 with <em>Dsvasa</em> promoter was introduced into the <em>Dstra</em> locus. Abnormal development of both the external genitalia and gonads was observed in G0 and G1 chromosomal female adults that expressed the male-specific <em>doublesex</em> (<em>dsx</em>) transcript. Interestingly, knocking out <em>Dstra</em> led to significantly reduced fertility in adults of corresponding sex and moderate transmission rates of the DsRed gene (63.54 %) were observed. Our results not only confirm the conserved function of the <em>Dstra</em> gene in sex determination, but also highlight the potential of sex conversion-based suppression gene-drive strategy targeting the <em>Dstra</em> gene in controlling of <em>D. suzukii</em> populations.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104406"},"PeriodicalIF":3.7,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145155183","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":"V-ATPase B mediates Bt Cry1Ac binding and toxicity in Grapholita molesta","authors":"Dandan Pan ,&nbsp;Jiayang Feng ,&nbsp;Jiacheng Ye,&nbsp;Xiaoyan Yang,&nbsp;Xiaoyan Zhang,&nbsp;Xiaofang Sha,&nbsp;Donghan Wang,&nbsp;Yanshen Fu,&nbsp;Chunxiao Men,&nbsp;Xiangqun Yuan,&nbsp;Yiping Li","doi":"10.1016/j.ibmb.2025.104407","DOIUrl":"10.1016/j.ibmb.2025.104407","url":null,"abstract":"<div><div><em>Grapholita molesta</em> is a worldwide pest. Cry1Ac is a significant alternative <em>Bacillus thuringiensis</em> (Bt) protein that exhibits substantial toxicity towards Lepidoptera insects. This investigation employed ligand blot, SDS-PAGE and liquid chromatography-tandem mass spectrometry (LC-MS/MS) techniques to identify the proteins potentially binding Cry1Ac on the peritrophic membrane (PM) of <em>G. molesta</em>. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis revealed the highest expression of <em>GmolV-ATPase B</em> in the midgut of 4th instar larvae. Following the ingestion of Cry1Ac protoxin by <em>G. molesta</em> larvae, a notable reduction in the expression level of <em>GmolV-ATPase B</em> was observed. The interaction between GmolV-ATPase B and activated Cry1Ac toxin was confirmed through ligand blot and homologous and heterologous competition experiments. Overexpression of GmolV-ATPase B in Sf9 cells led to an increase in Cry1Ac cytotoxicity, while RNAi targeting <em>GmolV-ATPase B</em> resulted in reduced mortality. In vivo bioassays demonstrated that the combined action of GmolV-ATPase B protein and Cry1Ac protoxin significantly enhanced the toxicity of Cry1Ac towards <em>G. molesta</em> larvae compared to Cry1Ac alone. These findings shed light on the binding of Cry1Ac to PM of <em>G. molesta</em> and its insecticidal mechanism, offering a valuable important reference for the development of biopesticides targeting midgut PM proteins.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"185 ","pages":"Article 104407"},"PeriodicalIF":3.7,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145172182","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}