Pub Date : 2025-12-02DOI: 10.1016/j.ibmb.2025.104456
Luke J. Pfannenstiel , Rachel H. Norris , Tobias Ziemke , Christophe Duplais , Nicolas Buchon , Jeffrey G. Scott
Nicotine is a plant-derived pyridine alkaloid with potent neurotoxic properties. A major pathway for detoxification of nicotine in mammals is via glucuronidation to produce nicotine N-glucuronide, but this process in insects remains poorly understood. Using mass spectrometry, we demonstrate that Drosophila melanogaster detoxifies nicotine through glycosylation, producing nicotine N-glycoside. Given that many new agrochemicals contain pyridine rings, we also investigated the metabolism of flonicamid and imidacloprid. We detected glycosylation of flonicamid, but not imidacloprid. A targeted RNAi screen across 21 UDP-glycosyltransferases (Ugts) identified Ugt35B1 as important for survival of nicotine exposure. CRISPR-based knockout of Ugt35B1 increases sensitivity to nicotine and flonicamid, but not to imidacloprid, nor to a structurally distinct neonicotinoid (thiamethoxam). Mass spectrometry of knockout and control flies confirms that Ugt35B1 glycosylates nicotine, its metabolite cotinine, and flonicamid. Together these findings establish Ugt35B1 as an important UGT mediating nicotine detoxification in adult D. melanogaster, revealing a previously uncharacterized insect glycosylation pathway with potential implications for herbivory, insecticide detoxification and toxicology.
{"title":"UGT35B1 is the principal enzyme mediating nicotine glycosylation in adult Drosophila melanogaster","authors":"Luke J. Pfannenstiel , Rachel H. Norris , Tobias Ziemke , Christophe Duplais , Nicolas Buchon , Jeffrey G. Scott","doi":"10.1016/j.ibmb.2025.104456","DOIUrl":"10.1016/j.ibmb.2025.104456","url":null,"abstract":"<div><div>Nicotine is a plant-derived pyridine alkaloid with potent neurotoxic properties. A major pathway for detoxification of nicotine in mammals is via glucuronidation to produce nicotine <em>N</em>-glucuronide, but this process in insects remains poorly understood. Using mass spectrometry, we demonstrate that <em>Drosophila melanogaster</em> detoxifies nicotine through glycosylation, producing nicotine <em>N</em>-glycoside. Given that many new agrochemicals contain pyridine rings, we also investigated the metabolism of flonicamid and imidacloprid. We detected glycosylation of flonicamid, but not imidacloprid. A targeted RNAi screen across 21 UDP-glycosyltransferases (<em>Ugt</em>s) identified <em>Ugt35B1</em> as important for survival of nicotine exposure. CRISPR-based knockout of <em>Ugt35B1</em> increases sensitivity to nicotine and flonicamid, but not to imidacloprid, nor to a structurally distinct neonicotinoid (thiamethoxam). Mass spectrometry of knockout and control flies confirms that <em>Ugt35B1</em> glycosylates nicotine, its metabolite cotinine, and flonicamid. Together these findings establish <em>Ugt35B1</em> as an important UGT mediating nicotine detoxification in adult <em>D. melanogaster</em>, revealing a previously uncharacterized insect glycosylation pathway with potential implications for herbivory, insecticide detoxification and toxicology.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"187 ","pages":"Article 104456"},"PeriodicalIF":3.7,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145675883","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}
Pub Date : 2025-12-02DOI: 10.1016/j.ibmb.2025.104457
Woo-Ram Park , Byungyoon Choi , Nanthini Sadasivam , Hui-Jin Bae , Sunmin Kim , Eunae Kim , Hueng-Sik Choi , In-Hong Jeong , Don-Kyu Kim
Insecticides are widely used in pest control; however, the increasing development of resistance in pests poses a significant global challenge. Nuclear receptors (NRs), ligand-dependent transcription factors, regulate the expression of genes involved in diverse metabolic processes, including development, detoxification, and innate immunity. However, the role of NRs in the detoxification processes of the brown planthopper, Nilaparvata lugens (N. lugens), remains poorly understood. Here, we show that estrogen-related receptor (NlERR) is a novel regulator of cytochrome P450 (CYP) genes for imidacloprid (IMD) resistance in N. lugens. Interestingly, the NlERR was significantly overexpressed in IMD-resistant strains. Genome-wide transcriptome analysis of NlERR-deficient N. lugens showed a positive correlation between the NlERR and the transcription of genes associated with detoxification metabolism. In addition, IMD treatment significantly increased the gene expression of phase I P450 enzymes in suspectable N. lugens, which was reversed by NlERR knockdown. Moreover, chromatin immunoprecipitation analysis confirmed that the NlERR directly binds to the ERR-response elements on the promoter of the CYP4CE1 gene. Consequently, NlERR knockdown in IMD-resistant strains significantly reduced the survival rate following IMD treatment and decreased expression of its target genes, CYP4CE1 and CYP6CW1. Finally, silencing either CYP4CE1 or CYP6CW1 in IMD-resistant strains significantly decreased the survival rate of the strains treated with IMD treatment. These findings establish NlERR as a key genetic factor for conferring IMD resistance in the N. lugens. Selectively controlling NlERR activity with a specific modulator will provide critical insights for developing new strategies to combat insecticide resistance in the N. lugens.
杀虫剂广泛应用于害虫防治；然而，害虫抗性的日益发展对全球构成了重大挑战。核受体（NRs），配体依赖性转录因子，调节参与多种代谢过程的基因表达，包括发育、解毒和先天免疫。然而，NRs在褐飞虱（Nilaparvata lugens， N. lugens）解毒过程中的作用仍然知之甚少。在这里，我们发现雌激素相关受体（NlERR）是一种新的细胞色素P450 （CYP）基因的调节剂，用于吡虫啉（IMD）抗性。有趣的是，NlERR在抗imd菌株中显著过表达。对NlERR缺失的N. lugens的全基因组转录组分析显示，NlERR与解毒代谢相关基因的转录呈正相关。此外，IMD处理显著增加了可疑N. lugens中I期P450酶的基因表达，这一现象被NlERR敲低逆转。此外，染色质免疫沉淀分析证实，NlERR直接与CYP4CE1基因启动子上的err应答元件结合。因此，耐IMD菌株的NlERR敲低显著降低了IMD治疗后的存活率，并降低了其靶基因CYP4CE1和CYP6CW1的表达。最后，沉默IMD耐药菌株中的CYP4CE1或CYP6CW1显著降低了IMD处理菌株的存活率。这些研究结果表明，NlERR是赋予N. lugens对IMD抗性的关键遗传因素。用一种特定的调节剂选择性地控制NlERR活性将为开发新的策略来对抗N. lugens的杀虫剂抗性提供重要的见解。
{"title":"Nuclear receptor ERR contributes to imidacloprid resistance by upregulating cytochrome P450 in Nilaparvata lugens","authors":"Woo-Ram Park , Byungyoon Choi , Nanthini Sadasivam , Hui-Jin Bae , Sunmin Kim , Eunae Kim , Hueng-Sik Choi , In-Hong Jeong , Don-Kyu Kim","doi":"10.1016/j.ibmb.2025.104457","DOIUrl":"10.1016/j.ibmb.2025.104457","url":null,"abstract":"<div><div>Insecticides are widely used in pest control; however, the increasing development of resistance in pests poses a significant global challenge. Nuclear receptors (NRs), ligand-dependent transcription factors, regulate the expression of genes involved in diverse metabolic processes, including development, detoxification, and innate immunity. However, the role of NRs in the detoxification processes of the brown planthopper, <em>Nilaparvata lugens</em> (<em>N. lugens</em>), remains poorly understood. Here, we show that estrogen-related receptor (NlERR) is a novel regulator of cytochrome P450 (CYP) genes for imidacloprid (IMD) resistance in <em>N. lugens</em>. Interestingly, the NlERR was significantly overexpressed in IMD-resistant strains. Genome-wide transcriptome analysis of NlERR-deficient <em>N. lugens</em> showed a positive correlation between the NlERR and the transcription of genes associated with detoxification metabolism. In addition, IMD treatment significantly increased the gene expression of phase I P450 enzymes in suspectable <em>N. lugens</em>, which was reversed by NlERR knockdown. Moreover, chromatin immunoprecipitation analysis confirmed that the NlERR directly binds to the ERR-response elements on the promoter of the CYP4CE1 gene. Consequently, NlERR knockdown in IMD-resistant strains significantly reduced the survival rate following IMD treatment and decreased expression of its target genes, CYP4CE1 and CYP6CW1. Finally, silencing either CYP4CE1 or CYP6CW1 in IMD-resistant strains significantly decreased the survival rate of the strains treated with IMD treatment. These findings establish NlERR as a key genetic factor for conferring IMD resistance in the <em>N. lugens</em>. Selectively controlling NlERR activity with a specific modulator will provide critical insights for developing new strategies to combat insecticide resistance in the <em>N. lugens</em>.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"187 ","pages":"Article 104457"},"PeriodicalIF":3.7,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145658832","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}
Pub Date : 2025-11-29DOI: 10.1016/j.ibmb.2025.104454
Shuai Wang , Lifei Qiu , Yashuai Wu , Zhenfang Li , Zhengwei Wang , Zhenhua Chen , Shanlin Liu , Xin Zhou
Temperature-induced variation in body coloration is a widespread form of phenotypic plasticity among insects, often mediated by the differential expression of pigmentation-related genes. Despite extensive documentation of this phenomenon, the molecular mechanisms by which environmental cues modulate pigmentation pathways remain largely unresolved. In this study, we investigated seasonal darkness plasticity in Apis cerana, the Eastern honey bee exhibiting a characteristic “yellow-black” abdominal pattern that responds dynamically to ambient temperature. Individuals reared under lower temperature conditions developed a markedly darker phenotype compared to those maintained at higher temperatures, highlighting a temperature-dependent shift in cuticular pigmentation. Exposure to low temperature also resulted in thicker adult cuticles and a concomitant reduction in water loss rate. The co-occurrence of darkened pigmentation and cuticle thickening corresponds with the colder and drier conditions characteristic of winter in the natural habitat of A. cerana. Furthermore, the Hedgehog (Hh) pathway was found to be enriched during early development stages. Notably, its target gene optomotor-blind (omb), a key temperature-sensitive regulator, was upregulated under decreased temperature, establishing the spatial arrangement of the black stripe at the posterior end of each abdominal tergite. Moreover, transcriptional suppression of omb induced upregulation of the pigmentation gene ebony, whose intrinsic thermally responsive expression directly enhances phenotypic plasticity in epidermal pigmentation, serving as a regulatory amplifier for environment-dependent melanin patterning. At the Pb (black eyes with an unpigmented body) in the mid-pupal period, temperature significantly influences 20-hydroxyecdysone (20E) titers. Integrated analyses combining in vivo 20E injection and LC-MS-based hormone quantification revealed that elevated developmental temperature upregulates 20E titers during the temperature-sensitive window, thereby suppressing the expression of pigmentation-related genes. These findings reveal a two-phase mechanism in which spatial patterning and hormonal signaling are temporally decoupled: omb acts early to establish the pigmentation boundary, while 20E acts later to adjust pigment intensity in a temperature-dependent manner. This layered control system allows A. cerana to fine-tune both the position and extent of abdominal pigmentation, providing a flexible strategy for thermal adaptation.
{"title":"Temperature-responsive spatial-temporal regulation underlies phenotypic plasticity of body pigmentation in Eastern honey bee","authors":"Shuai Wang , Lifei Qiu , Yashuai Wu , Zhenfang Li , Zhengwei Wang , Zhenhua Chen , Shanlin Liu , Xin Zhou","doi":"10.1016/j.ibmb.2025.104454","DOIUrl":"10.1016/j.ibmb.2025.104454","url":null,"abstract":"<div><div>Temperature-induced variation in body coloration is a widespread form of phenotypic plasticity among insects, often mediated by the differential expression of pigmentation-related genes. Despite extensive documentation of this phenomenon, the molecular mechanisms by which environmental cues modulate pigmentation pathways remain largely unresolved. In this study, we investigated seasonal darkness plasticity in <em>Apis cerana</em>, the Eastern honey bee exhibiting a characteristic “yellow-black” abdominal pattern that responds dynamically to ambient temperature. Individuals reared under lower temperature conditions developed a markedly darker phenotype compared to those maintained at higher temperatures, highlighting a temperature-dependent shift in cuticular pigmentation. Exposure to low temperature also resulted in thicker adult cuticles and a concomitant reduction in water loss rate. The co-occurrence of darkened pigmentation and cuticle thickening corresponds with the colder and drier conditions characteristic of winter in the natural habitat of <em>A. cerana</em>. Furthermore<em>,</em> the Hedgehog (Hh) pathway was found to be enriched during early development stages. Notably, its target gene <em>optomotor-blind</em> (<em>omb</em>), a key temperature-sensitive regulator, was upregulated under decreased temperature, establishing the spatial arrangement of the black stripe at the posterior end of each abdominal tergite. Moreover, transcriptional suppression of <em>omb</em> induced upregulation of the pigmentation gene <em>ebony</em>, whose intrinsic thermally responsive expression directly enhances phenotypic plasticity in epidermal pigmentation, serving as a regulatory amplifier for environment-dependent melanin patterning. At the Pb (black eyes with an unpigmented body) in the mid-pupal period, temperature significantly influences 20-hydroxyecdysone (20E) titers. Integrated analyses combining <em>in vivo</em> 20E injection and LC-MS-based hormone quantification revealed that elevated developmental temperature upregulates 20E titers during the temperature-sensitive window, thereby suppressing the expression of pigmentation-related genes. These findings reveal a two-phase mechanism in which spatial patterning and hormonal signaling are temporally decoupled: <em>omb</em> acts early to establish the pigmentation boundary, while 20E acts later to adjust pigment intensity in a temperature-dependent manner. This layered control system allows <em>A. cerana</em> to fine-tune both the position and extent of abdominal pigmentation, providing a flexible strategy for thermal adaptation.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104454"},"PeriodicalIF":3.7,"publicationDate":"2025-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145652935","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}
Pub Date : 2025-11-21DOI: 10.1016/j.ibmb.2025.104453
Mengqing Deng, Xiyue Xu, Zhiming Yang, Kai Lu
The escalating problem of insecticide resistance in agricultural pests highlights the need to identify non-canonical resistance mechanisms. In this study, comparative transcriptional analysis identified five chemosensory protein (CSP) genes significantly overexpressed in a chlorpyrifos (CPF)-resistant strain of Nilaparvata lugens. RNA interference and heterologous expression confirmed their contribution to resistance, with CSP15, exhibiting 42.98-fold overexpression, identified as a key mediator. CSP15 knockdown increased CPF susceptibility 1.76-fold, while its bacterial expression raised tolerance 5.47-fold. Competitive fluorescence binding assays showed that CSP15 binds CPF (Ki = 3.32 μM) with higher affinity than its toxic metabolite chlorpyrifos-oxon (CPO; Ki = 6.00 μM). Structural modeling revealed a CPF-binding pocket mediated by Arg84 and Lys118. CPO binding utilized four hydrogen bonds with Lys28; the K28A mutation induced structural rearrangement through neo-formed interactions with Arg84 and Lys118, reducing the binding free energy by −40.95 kcal mol−1. Mutagenesis revealed that R84A/K118A double mutants lost 62.85–64.37 % of CPF-binding capacity, whereas the K28A variant increased CPO affinity by 22.65 %. In vivo metabolic profiling indicated that CSP15 knockdown promoted CPF degradation and CPO accumulation, supporting its role in suppressing bioactivation. Yeast two-hybrid assays confirmed a direct interaction between CSP15 and CYP6BD12, the P450 enzyme responsible for CPF bioactivation. Additionally, CSP15 expression was regulated by the transcription factor Lim1β through two conserved promoter cis-elements. Together, these findings reveal a novel dual resistance mechanism wherein CSP15 acts synergistically by sequestering CPF and binding CYP6BD12, thereby effectively suppressing bioactivation and identifying promising targets for pest management.
{"title":"CSP15 attenuates chlorpyrifos toxicity through sequestration and bioactivation suppression in Nilaparvata lugens","authors":"Mengqing Deng, Xiyue Xu, Zhiming Yang, Kai Lu","doi":"10.1016/j.ibmb.2025.104453","DOIUrl":"10.1016/j.ibmb.2025.104453","url":null,"abstract":"<div><div>The escalating problem of insecticide resistance in agricultural pests highlights the need to identify non-canonical resistance mechanisms. In this study, comparative transcriptional analysis identified five chemosensory protein (CSP) genes significantly overexpressed in a chlorpyrifos (CPF)-resistant strain of <em>Nilaparvata lugens</em>. RNA interference and heterologous expression confirmed their contribution to resistance, with <em>CSP15</em>, exhibiting 42.98-fold overexpression, identified as a key mediator. <em>CSP15</em> knockdown increased CPF susceptibility 1.76-fold, while its bacterial expression raised tolerance 5.47-fold. Competitive fluorescence binding assays showed that CSP15 binds CPF (<em>K</em><sub>i</sub> = 3.32 μM) with higher affinity than its toxic metabolite chlorpyrifos-oxon (CPO; <em>K</em><sub>i</sub> = 6.00 μM). Structural modeling revealed a CPF-binding pocket mediated by Arg84 and Lys118. CPO binding utilized four hydrogen bonds with Lys28; the K28A mutation induced structural rearrangement through neo-formed interactions with Arg84 and Lys118, reducing the binding free energy by −40.95 kcal mol<sup>−1</sup>. Mutagenesis revealed that R84A/K118A double mutants lost 62.85–64.37 % of CPF-binding capacity, whereas the K28A variant increased CPO affinity by 22.65 %. <em>In vivo</em> metabolic profiling indicated that <em>CSP15</em> knockdown promoted CPF degradation and CPO accumulation, supporting its role in suppressing bioactivation. Yeast two-hybrid assays confirmed a direct interaction between CSP15 and CYP6BD12, the P450 enzyme responsible for CPF bioactivation. Additionally, <em>CSP15</em> expression was regulated by the transcription factor Lim1β through two conserved promoter <em>cis</em>-elements. Together, these findings reveal a novel dual resistance mechanism wherein CSP15 acts synergistically by sequestering CPF and binding CYP6BD12, thereby effectively suppressing bioactivation and identifying promising targets for pest management.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104453"},"PeriodicalIF":3.7,"publicationDate":"2025-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145577095","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}
Pub Date : 2025-11-20DOI: 10.1016/j.ibmb.2025.104452
Jiarui Tao , Xiao Wang , Changhong Xia , Xingyuan Wang , Yifan Zhang , Jianan Liu , Juhong Zhang , Jun Wang , Jinghui Xi , Shang Wang
The soil-dwelling grubs of Holotrichia parallela inflict substantial damage to the root systems of key cereal crops, including maize and peanuts, thereby posing a serious threat to agricultural productivity. While the olfactory mechanisms through which above-ground insects utilize odorant receptors (ORs) and odorant co-receptors (Orco) to detect plant volatiles for host localization are well established, the role of OR-mediated chemoreception in the host-seeking behaviors of underground pests remains poorly understood. To address this gap, we systematically compared the volatile compounds released by rhizospheric and aerial plant tissues. Through antennal transcriptome analysis and sequence structure prediction, we identified an odorant receptor with biased olfactory expression—HparOR48. Tissue expression analysis revealed that HparOR48 is predominantly expressed in the larval antennae across developmental stages. The functional characterization of HparOR48 was performed using the Xenopus laevis oocyte expression system, which demonstrated its broad responsiveness to phthalic acid, a root-emitted acidic volatile. Electroantennogram (EAG) assays further confirmed that phthalic acid elicited significant electrophysiological responses in the larval antennae. Moreover, behavioral assays utilizing a four-arm olfactometer indicated that phthalic acid acts as an attractant for the larvae. Our findings provide clear evidence that H. parallela larvae rely on olfactory recognition of rhizosphere-specific acidic volatiles to guide host-seeking behavior, thereby extending our understanding of chemical interactions between soil-dwelling pests and their host plants.
{"title":"Herbivore-induced plant root volatiles phthalic acid mediate host selection for Holotrichia parallela larva","authors":"Jiarui Tao , Xiao Wang , Changhong Xia , Xingyuan Wang , Yifan Zhang , Jianan Liu , Juhong Zhang , Jun Wang , Jinghui Xi , Shang Wang","doi":"10.1016/j.ibmb.2025.104452","DOIUrl":"10.1016/j.ibmb.2025.104452","url":null,"abstract":"<div><div>The soil-dwelling grubs of <em>Holotrichia parallela</em> inflict substantial damage to the root systems of key cereal crops, including maize and peanuts, thereby posing a serious threat to agricultural productivity. While the olfactory mechanisms through which above-ground insects utilize odorant receptors (ORs) and odorant co-receptors (Orco) to detect plant volatiles for host localization are well established, the role of OR-mediated chemoreception in the host-seeking behaviors of underground pests remains poorly understood. To address this gap, we systematically compared the volatile compounds released by rhizospheric and aerial plant tissues. Through antennal transcriptome analysis and sequence structure prediction, we identified an odorant receptor with biased olfactory expression—<em>HparOR48</em>. Tissue expression analysis revealed that <em>HparOR48</em> is predominantly expressed in the larval antennae across developmental stages. The functional characterization of <em>HparOR48</em> was performed using the <em>Xenopus laevis oocyte</em> expression system, which demonstrated its broad responsiveness to phthalic acid, a root-emitted acidic volatile. Electroantennogram (EAG) assays further confirmed that phthalic acid elicited significant electrophysiological responses in the larval antennae. Moreover, behavioral assays utilizing a four-arm olfactometer indicated that phthalic acid acts as an attractant for the larvae. Our findings provide clear evidence that H. parallela larvae rely on olfactory recognition of rhizosphere-specific acidic volatiles to guide host-seeking behavior, thereby extending our understanding of chemical interactions between soil-dwelling pests and their host plants.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104452"},"PeriodicalIF":3.7,"publicationDate":"2025-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145577096","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}
Pub Date : 2025-11-20DOI: 10.1016/j.ibmb.2025.104451
Jiangjie Li , Yimeng Chen , Jing Wang, Clauvis NT. Taning, Kristof De Schutter
Sex determination is a fundamental process during early embryonic development that shapes the morphological, physiological, and behavioral sexual dimorphism of an organism. Insects, the most diverse group of animals on earth, have evolved a wide range of sex determination systems. This study investigates the sex determination and differentiation in the Colorado potato beetle (CPB), a species with an X0 sex chromosome system, by identifying key regulatory genes involved in these processes. Our findings reveal that the genes Sex-lethal (Sxl), transformer-2 (tra2), doublesex (dsx), and fruitless (fru) are highly conserved and all contribute to fertility. Among them, dsx and tra2 emerge as key genetic switches of sex differentiation in CPB: Disruption of dsx leads to feminization of male traits and abnormal female development. Additionally, tra2 appears to regulate the alternative splicing of dsx, promoting the production of the female-specific transcripts. This study highlights the pivotal roles of dsx and tra2 in CPB sex determination and provides valuable insights into the evolution and diversity of sex determination mechanisms in insects.
{"title":"Master genetic switches controlling sexual development in Leptinotarsa decemlineata","authors":"Jiangjie Li , Yimeng Chen , Jing Wang, Clauvis NT. Taning, Kristof De Schutter","doi":"10.1016/j.ibmb.2025.104451","DOIUrl":"10.1016/j.ibmb.2025.104451","url":null,"abstract":"<div><div>Sex determination is a fundamental process during early embryonic development that shapes the morphological, physiological, and behavioral sexual dimorphism of an organism. Insects, the most diverse group of animals on earth, have evolved a wide range of sex determination systems. This study investigates the sex determination and differentiation in the Colorado potato beetle (CPB), a species with an X0 sex chromosome system, by identifying key regulatory genes involved in these processes. Our findings reveal that the genes <em>Sex-lethal</em> (<em>Sxl</em>), <em>transformer-2</em> (<em>tra2</em>), <em>doublesex</em> (<em>dsx</em>), and <em>fruitless</em> (<em>fru</em>) are highly conserved and all contribute to fertility. Among them, <em>dsx</em> and <em>tra2</em> emerge as key genetic switches of sex differentiation in CPB: Disruption of <em>dsx</em> leads to feminization of male traits and abnormal female development. Additionally, <em>tra2</em> appears to regulate the alternative splicing of <em>dsx</em>, promoting the production of the female-specific transcripts. This study highlights the pivotal roles of <em>dsx</em> and <em>tra2</em> in CPB sex determination and provides valuable insights into the evolution and diversity of sex determination mechanisms in insects.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104451"},"PeriodicalIF":3.7,"publicationDate":"2025-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145577126","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}
Pub Date : 2025-11-19DOI: 10.1016/j.ibmb.2025.104450
Wen-qing Lai , Zhan-peng Lu , Qiang Zhang , Zuo-min Shao , Han Gao , Xia Sun , Sheng Qin , Xue-yang Wang , Mu-wang Li
Insect reproductive capacity is a key determinant of population fitness, though its regulatory mechanisms remain limited. Here, we identified an op50 silkworm mutant (derived from p50 strain) exhibiting female-specific reproductive defects, which provided an important model for studying insect reproductive regulation mechanism. We found that Bombyx mori xanthine dehydrogenase 1 (BmXDH1) in op50 contained a single-base insertion causing premature termination, deleting 154 C-terminal amino acids and losing catalytic activity. ΔBmXDH1 mutants recapitulated the op50 phenotype with 50 % reduced oviposition and 90 % lower hatchability, attributed to severe structural impairments of the eggshell surface and egg surface pore. In addition, female reproductive defects initiated at the pupal stage, characterized by oviduct malformation and impaired oviposition. This phenotype was associated with excessive accumulation of hypoxanthine, leading to ovarian and embryonic developmental disruption, including extracellular matrix remodeling, cellular signaling pathways, and oxidative stress regulation. The results uncovered how BmXDH1 regulates hypoxanthine metabolism to control silkworm reproduction, advancing understanding of insect reproductive networks and providing targets for pest control/economic insect breeding.
{"title":"Hypoxanthine accumulation caused by mutations of Bombyx mori xanthine dehydrogenase triggers female sterility","authors":"Wen-qing Lai , Zhan-peng Lu , Qiang Zhang , Zuo-min Shao , Han Gao , Xia Sun , Sheng Qin , Xue-yang Wang , Mu-wang Li","doi":"10.1016/j.ibmb.2025.104450","DOIUrl":"10.1016/j.ibmb.2025.104450","url":null,"abstract":"<div><div>Insect reproductive capacity is a key determinant of population fitness, though its regulatory mechanisms remain limited. Here, we identified an op50 silkworm mutant (derived from p50 strain) exhibiting female-specific reproductive defects, which provided an important model for studying insect reproductive regulation mechanism. We found that <em>Bombyx mori xanthine dehydrogenase 1</em> (<em>BmXDH1</em>) in op50 contained a single-base insertion causing premature termination, deleting 154 C-terminal amino acids and losing catalytic activity. <em>ΔBmXDH1</em> mutants recapitulated the op50 phenotype with 50 % reduced oviposition and 90 % lower hatchability, attributed to severe structural impairments of the eggshell surface and egg surface pore. In addition, female reproductive defects initiated at the pupal stage, characterized by oviduct malformation and impaired oviposition. This phenotype was associated with excessive accumulation of hypoxanthine, leading to ovarian and embryonic developmental disruption, including extracellular matrix remodeling, cellular signaling pathways, and oxidative stress regulation. The results uncovered how <em>BmXDH1</em> regulates hypoxanthine metabolism to control silkworm reproduction, advancing understanding of insect reproductive networks and providing targets for pest control/economic insect breeding.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104450"},"PeriodicalIF":3.7,"publicationDate":"2025-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145562280","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}
Pub Date : 2025-11-14DOI: 10.1016/j.ibmb.2025.104449
Na Zhang , Xuemin Zhou , Xinhao Jiao , Zi Liang , Wenwen Jiang , Shuyi Liu , Ping Wu
Simple, accurate, sensitive, and rapid pathogen diagnosis is crucial for effective control of silkworm diseases. Although CRISPR-based nucleic acid detection systems show great potential for on-site detection of silkworm pathogens, their practicality is hindered by complex workflows and reagent-storage constraints. To address these limitations and enhance field suitability, we developed a DNA extraction-free one-pot RPA-CRISPR/Cas12a (DEORC) system and a dual-gene assay for detecting Bombyx mori nucleopolyhedrovirus (BmNPV) and Nosema bombycis using a handheld device. The DEORC assay detects BmNPV in hemolymph samples as early as 6 h post-infection (hpi) and N. bombycis at 103 spores/mL in spore suspensions. The entire process from sampling to visual readout is completed in approximately 70 min without requiring sophisticated equipment. To further enable off-grid deployment, we lyophilized the Cas12a detection reagents using 1 M betaine as a lyoprotectant, which retained measurable activity for at least one month at 4 °C under our test conditions, facilitating short-term refrigerated transport and field storage. Additionally, the dual-gene assay detects 103 copies/μL of a double-reference plasmid and simultaneously detects both BmNPV and N. bombycis in a single tube from midgut samples at 48 hpi; when combined with extraction-free techniques, it enables simultaneous detection of both pathogens in hemolymph samples at 72 hpi. Collectively, these advancements provide sensitive and portable tools for on-site sericulture disease management, offering faster and more practical workflows than two-step single-gene and traditional approaches.
{"title":"Field-deployable CRISPR-Dx for BmNPV and Nosema bombycis: DNA-extraction-free one-pot RPA-Cas12a and Cas12a/Cas13a dual-gene assays with handheld devices","authors":"Na Zhang , Xuemin Zhou , Xinhao Jiao , Zi Liang , Wenwen Jiang , Shuyi Liu , Ping Wu","doi":"10.1016/j.ibmb.2025.104449","DOIUrl":"10.1016/j.ibmb.2025.104449","url":null,"abstract":"<div><div>Simple, accurate, sensitive, and rapid pathogen diagnosis is crucial for effective control of silkworm diseases. Although CRISPR-based nucleic acid detection systems show great potential for on-site detection of silkworm pathogens, their practicality is hindered by complex workflows and reagent-storage constraints. To address these limitations and enhance field suitability, we developed a DNA extraction-free one-pot RPA-CRISPR/Cas12a (DEORC) system and a dual-gene assay for detecting <em>Bombyx mori nucleopolyhedrovirus</em> (BmNPV) and <em>Nosema bombycis</em> using a handheld device. The DEORC assay detects BmNPV in hemolymph samples as early as 6 h post-infection (hpi) and <em>N. bombycis</em> at 10<sup>3</sup> spores/mL in spore suspensions. The entire process from sampling to visual readout is completed in approximately 70 min without requiring sophisticated equipment. To further enable off-grid deployment, we lyophilized the Cas12a detection reagents using 1 M betaine as a lyoprotectant, which retained measurable activity for at least one month at 4 °C under our test conditions, facilitating short-term refrigerated transport and field storage. Additionally, the dual-gene assay detects 10<sup>3</sup> copies/μL of a double-reference plasmid and simultaneously detects both BmNPV and <em>N. bombycis</em> in a single tube from midgut samples at 48 hpi; when combined with extraction-free techniques, it enables simultaneous detection of both pathogens in hemolymph samples at 72 hpi. Collectively, these advancements provide sensitive and portable tools for on-site sericulture disease management, offering faster and more practical workflows than two-step single-gene and traditional approaches.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104449"},"PeriodicalIF":3.7,"publicationDate":"2025-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145533966","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}
Pub Date : 2025-11-13DOI: 10.1016/j.ibmb.2025.104447
Peng Zeng , Xiaodong Li , Xuan Jin , Qien Zhong , Fareed Uddin Memon , Youliang Pan , Yang Xiao , Kang Li , Ling Tian
Lysosomal Acid Lipase (LAL) is the key enzyme responsible for hydrolyzing cholesteryl esters and triglycerides within lysosomes. Its dysfunction is linked to various metabolic disorders in humans. However, its biological functions display tissue-specific heterogeneity, and the roles of its homologs in invertebrates remain largely unexplored. Herein, we demonstrated the lysosomal localization of acid lipase-1 (BmAL1) in Bombyx mori through fluorescent protein observation and Co-immunoprecipitation with the lysosomal membrane glycoprotein BmLAMP1. BmAL1 overexpression promoted lysosomal acidification, enhanced autophagy, and reduced lipid droplet (LD) formation after oleic acid or palmitic acid treatment. Conversely, BmAL1 knockout or knockdown inhibited lysosomal acidification, autophagic flux, and lipid degradation. Subsequently, immunoprecipitation coupled with LC-MS analysis revealed that the BmAL1-interacting proteins were mainly associated with metabolic and human-disease pathways. Notably, functional disruption of BmAL1-interacting protein BmHSP83/BmHSP90 (heat shock protein) compromised BmAL1-mediated lysosomal acidification, autophagy induction, lipid metabolism, and the physical interaction between BmAL1 and BmLAMP1. These data fill a critical gaps in LAL research in invertebrates, and highlight potential targets for the utilization of beneficial insects and the development of pest control strategies.
{"title":"Lysosomal acid lipase-1 functions in autophagy occurrence and lipid homeostasis dependent of heat shock protein 83 in insects","authors":"Peng Zeng , Xiaodong Li , Xuan Jin , Qien Zhong , Fareed Uddin Memon , Youliang Pan , Yang Xiao , Kang Li , Ling Tian","doi":"10.1016/j.ibmb.2025.104447","DOIUrl":"10.1016/j.ibmb.2025.104447","url":null,"abstract":"<div><div>Lysosomal Acid Lipase (LAL) is the key enzyme responsible for hydrolyzing cholesteryl esters and triglycerides within lysosomes. Its dysfunction is linked to various metabolic disorders in humans. However, its biological functions display tissue-specific heterogeneity, and the roles of its homologs in invertebrates remain largely unexplored. Herein, we demonstrated the lysosomal localization of acid lipase-1 (BmAL1) in <em>Bombyx mori</em> through fluorescent protein observation and Co-immunoprecipitation with the lysosomal membrane glycoprotein BmLAMP1. <em>BmAL1</em> overexpression promoted lysosomal acidification, enhanced autophagy, and reduced lipid droplet (LD) formation after oleic acid or palmitic acid treatment. Conversely, <em>BmAL1</em> knockout or knockdown inhibited lysosomal acidification, autophagic flux, and lipid degradation. Subsequently, immunoprecipitation coupled with LC-MS analysis revealed that the BmAL1-interacting proteins were mainly associated with metabolic and human-disease pathways. Notably, functional disruption of BmAL1-interacting protein BmHSP83/BmHSP90 (heat shock protein) compromised BmAL1-mediated lysosomal acidification, autophagy induction, lipid metabolism, and the physical interaction between BmAL1 and BmLAMP1. These data fill a critical gaps in LAL research in invertebrates, and highlight potential targets for the utilization of beneficial insects and the development of pest control strategies.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104447"},"PeriodicalIF":3.7,"publicationDate":"2025-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145527177","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}
Pub Date : 2025-11-13DOI: 10.1016/j.ibmb.2025.104446
Chen Yang , Yu Zhu He , Yu Xiang Wang , Xing Chao Yan , Xu Han , Zhi Jiang Zeng , Xu Jiang He
Honeybee caste differentiation is a prime example of phenotypic plasticity in eusocial insects and serves as a valuable model for epigenetic studies. Various epigenetic modifications such as non-coding RNAs and histone modifications, have been implicated in caste differentiation. However, the role of some epigenetic modifications such as DNA methylation still remains controversial. Similarly, mRNA m6A modification has been suggested to participate in honeybee caste differentiation. Here we used Oxford Nanopore technology to compare m6A modifications in 2d, 4d, and 6d queen and worker larvae. Our data indicated that m6A modification is unlikely to exert a direct role in the caste differentiation at the developmental stages examed in this study. We identified 363, 1074, and 1000 differentially expressed transcripts (DETs) in 2d, 4d, and 6d queen-worker larval comparisons, respectively, 74 of which were mapped in seven key KEGG pathways (e.g., mTOR, Notch, FoxO, MAPK, Wnt, Hippo, Hedgehog) associated with honeybee caste differentiation. In contrast, only 16, 39, and 11 differentially m6A-modified transcripts (m6A-DMTs) were found in the 2d, 4d and 6d comparisons, with very few m6A-DMTs involved in key pathways for caste differentiation. Notably, no correlation was observed between m6A modification levels and transcript expression, including DETs and m6A-DMTs. The motifs of m6A modifications remained unchanged across all three developmental time points. When the original samples were randomly regrouped, the number of m6A-DMTs did not change significantly, but the DETs related to caste differentiation were dramatically altered. Therefore, the functional relevance of m6A in honeybee caste differentiation is still under debate and requires more comprehensive studies.
{"title":"Limited association of m6A RNA modification with post-transcriptional regulation during honeybee caste differentiation","authors":"Chen Yang , Yu Zhu He , Yu Xiang Wang , Xing Chao Yan , Xu Han , Zhi Jiang Zeng , Xu Jiang He","doi":"10.1016/j.ibmb.2025.104446","DOIUrl":"10.1016/j.ibmb.2025.104446","url":null,"abstract":"<div><div>Honeybee caste differentiation is a prime example of phenotypic plasticity in eusocial insects and serves as a valuable model for epigenetic studies. Various epigenetic modifications such as non-coding RNAs and histone modifications, have been implicated in caste differentiation. However, the role of some epigenetic modifications such as DNA methylation still remains controversial. Similarly, mRNA m<sup>6</sup>A modification has been suggested to participate in honeybee caste differentiation. Here we used Oxford Nanopore technology to compare m<sup>6</sup>A modifications in 2d, 4d, and 6d queen and worker larvae. Our data indicated that m<sup>6</sup>A modification is unlikely to exert a direct role in the caste differentiation at the developmental stages examed in this study. We identified 363, 1074, and 1000 differentially expressed transcripts (DETs) in 2d, 4d, and 6d queen-worker larval comparisons, respectively, 74 of which were mapped in seven key KEGG pathways (e.g., mTOR, Notch, FoxO, MAPK, Wnt, Hippo, Hedgehog) associated with honeybee caste differentiation. In contrast, only 16, 39, and 11 differentially m<sup>6</sup>A-modified transcripts (m<sup>6</sup>A-DMTs) were found in the 2d, 4d and 6d comparisons, with very few m<sup>6</sup>A-DMTs involved in key pathways for caste differentiation. Notably, no correlation was observed between m<sup>6</sup>A modification levels and transcript expression, including DETs and m<sup>6</sup>A-DMTs. The motifs of m<sup>6</sup>A modifications remained unchanged across all three developmental time points. When the original samples were randomly regrouped, the number of m<sup>6</sup>A-DMTs did not change significantly, but the DETs related to caste differentiation were dramatically altered. Therefore, the functional relevance of m<sup>6</sup>A in honeybee caste differentiation is still under debate and requires more comprehensive studies.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104446"},"PeriodicalIF":3.7,"publicationDate":"2025-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145522484","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}
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