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}
Nectar serves as the primary carbohydrate source for honey bees, sustaining both individual and colony-wide energy demands. Although nectar from bee-pollinated flowers is rich in sucrose, a preferred sugar for honey bees, the neural mechanisms regulating sucrose intake remain poorly understood. Here, we demonstrated that the leucokinin (LK) neuropeptide and its receptor (LKR) suppress sucrose consumption in worker bees of Apis cerana, as shown through LK supplementation and Lkr RNAi. Gene silencing results revealed that this signaling pathway is inhibited by Lkrial (Lkr Intronic Antisense LncRNA), a novel long non-coding RNA (lncRNA) transcribed antisense to the Lkr locus. Furthermore, by using quantitative PCR and RNAi, we showed that Lk/Lkr signaling cascade activates the mechanosensory ion channel Piezo, which acts downstream to suppresses sucrose intake. Collectively, our findings uncover a previously unrecognized regulatory circuit that is essential for the neural control of sucrose consumption and energy acquisition in A. cerana.
{"title":"The leucokinin pathway regulates honey bee sugar consumption via Piezo","authors":"Zhenfang Li, Chengfeng Yang, Yashuai Wu, Xinyue Zhang, Xin Zhou, Shiqi Luo","doi":"10.1016/j.ibmb.2025.104448","DOIUrl":"10.1016/j.ibmb.2025.104448","url":null,"abstract":"<div><div>Nectar serves as the primary carbohydrate source for honey bees, sustaining both individual and colony-wide energy demands. Although nectar from bee-pollinated flowers is rich in sucrose, a preferred sugar for honey bees, the neural mechanisms regulating sucrose intake remain poorly understood. Here, we demonstrated that the leucokinin (LK) neuropeptide and its receptor (LKR) suppress sucrose consumption in worker bees of <em>Apis cerana</em>, as shown through LK supplementation and <em>Lkr</em> RNAi. Gene silencing results revealed that this signaling pathway is inhibited by <em>Lkrial</em> (<em>Lkr</em> Intronic Antisense LncRNA), a novel long non-coding RNA (lncRNA) transcribed antisense to the <em>Lkr</em> locus. Furthermore, by using quantitative PCR and RNAi, we showed that <em>Lk/Lkr</em> signaling cascade activates the mechanosensory ion channel <em>Piezo</em>, which acts downstream to suppresses sucrose intake. Collectively, our findings uncover a previously unrecognized regulatory circuit that is essential for the neural control of sucrose consumption and energy acquisition in <em>A. cerana</em>.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104448"},"PeriodicalIF":3.7,"publicationDate":"2025-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145511432","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 : 2025-11-08DOI: 10.1016/j.ibmb.2025.104444
Xue Kong , Dandan Li , Minghui Xu , Shaofeng Zhong , Shaoye Liu , Jeremiah Joe Kabissa , Yongyu Xu , Zhenzhen Chen
Reproductive diapause in insects is a crucial seasonal adaptation, predominantly regulated by photoperiod, with the circadian clock providing the internal time reference to measure day or night length. Despite its importance, the role of core circadian clock genes in photoperiodic responses and reproductive diapause induction remains poorly understood. The green lacewing, Chrysoperla nipponensis, an important predatory natural enemy, undergoes reproductive diapause under short-day (SD) conditions. This study aimed to investigate the role of circadian clock gene period (per) in photoperiod-mediated diapause of C. nipponensis. Firstly, we identified and molecularly characterized per in C. nipponensis. The encoded PER protein contains conserved domains (PAS, PAC, Period_C) and clusters phylogenetically with neuropteran and coleopteran orthologs. Next, we examined the spatiotemporal expression pattern of per, revealing that its expression was significantly influenced by photoperiod. The daily expression pattern of per indicates that it measures night length. Functional RNAi assays demonstrated that per knockdown under diapause-inducing (short-day) conditions upregulated other core clock genes (timeless1, cycle, clock), abolished diapause phenotypes (ovarian arrest, lipid accumulation), and triggered precocious reproduction. Furthermore, transcriptional analysis revealed that per silencing upregulates the core circadian clock gene cry1 as well as 20-hydroxyecdysone (20E) signaling genes, and reprograms lipid synthesis and metabolism genes under short-day conditions in C. nipponensis. Overall, these findings establish per as a critical genetic switch linking circadian timing to diapause induction in C. nipponensis, offering new insights into the molecular mechanisms of circadian regulation in seasonal adaptations of natural enemies, with potential applications in biological control of agricultural pests.
{"title":"Impact of the photoperiod-responsive circadian clock gene period on reproductive diapause in Chrysoperla nipponensis (Okamoto)","authors":"Xue Kong , Dandan Li , Minghui Xu , Shaofeng Zhong , Shaoye Liu , Jeremiah Joe Kabissa , Yongyu Xu , Zhenzhen Chen","doi":"10.1016/j.ibmb.2025.104444","DOIUrl":"10.1016/j.ibmb.2025.104444","url":null,"abstract":"<div><div>Reproductive diapause in insects is a crucial seasonal adaptation, predominantly regulated by photoperiod, with the circadian clock providing the internal time reference to measure day or night length. Despite its importance, the role of core circadian clock genes in photoperiodic responses and reproductive diapause induction remains poorly understood. The green lacewing, <em>Chrysoperla nipponensis</em>, an important predatory natural enemy, undergoes reproductive diapause under short-day (SD) conditions. This study aimed to investigate the role of circadian clock gene <em>period</em> (<em>per</em>) in photoperiod-mediated diapause of <em>C. nipponensis</em>. Firstly, we identified and molecularly characterized <em>per</em> in <em>C. nipponensis</em>. The encoded PER protein contains conserved domains (PAS, PAC, Period_C) and clusters phylogenetically with neuropteran and coleopteran orthologs. Next, we examined the spatiotemporal expression pattern of <em>per</em>, revealing that its expression was significantly influenced by photoperiod. The daily expression pattern of <em>per</em> indicates that it measures night length. Functional RNAi assays demonstrated that <em>per</em> knockdown under diapause-inducing (short-day) conditions upregulated other core clock genes (<em>timeless1</em>, <em>cycle</em>, <em>clock</em>), abolished diapause phenotypes (ovarian arrest, lipid accumulation), and triggered precocious reproduction. Furthermore, transcriptional analysis revealed that <em>per</em> silencing upregulates the core circadian clock gene <em>cry1</em> as well as 20-hydroxyecdysone (20E) signaling genes, and reprograms lipid synthesis and metabolism genes under short-day conditions in <em>C. nipponensis</em>. Overall, these findings establish <em>per</em> as a critical genetic switch linking circadian timing to diapause induction in <em>C. nipponensis</em>, offering new insights into the molecular mechanisms of circadian regulation in seasonal adaptations of natural enemies, with potential applications in biological control of agricultural pests.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104444"},"PeriodicalIF":3.7,"publicationDate":"2025-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145487347","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 : 2025-11-08DOI: 10.1016/j.ibmb.2025.104445
Mengqing Deng, Xiyue Xu, Zhiming Yang, Kai Lu
Insecticide resistance poses a critical challenge to global agricultural sustainability. While metabolic detoxification and target-site mutations are well-characterized resistance mechanisms, the role of micronutrient homeostasis remains understudied. This study reveals that biotin deficiency in Nilaparvata lugens drives imidacloprid resistance through a multi-tiered regulatory network coordinating chemosensory protein (CSP) dynamics. Biotin deficiency enhances CSP-mediated insecticide sequestration via high-affinity binding to CSP2, CSP4, CSP7, and CSP15, which are overexpressed in resistant strains. RNA interference and dual-luciferase assays demonstrate that the aryl hydrocarbon receptor and its nuclear translocator (AhR/ARNT) transcriptionally activate CSP2 and CSP15, with their knockdown partially restoring insecticide susceptibility. Furthermore, biotin deficiency activates reactive oxygen species (ROS)/cap ‘n’ collar C (CncC) signaling, elevating AhR/ARNT expression through transcriptional reprogramming. Yeast three-hybrid assays identify a post-translational regulatory layer, wherein biotin directly inhibits AhR–ARNT heterodimerization. Field-evolved resistant populations recapitulate this mechanism, exhibiting conserved molecular signatures including biotin deficiency, ROS/CncC pathway activation, and AhR/ARNT-CSP overexpression correlated with resistance intensity. These findings establish a unified model wherein biotin scarcity reprograms xenobiotic defense through three synergistic mechanisms: enhanced CSP–insecticide binding, transcriptional amplification via ROS/CncC-AhR/ARNT signaling, and post-translational optimization of transcriptional complexes. The operational conservation of this pathway across laboratory and ecological contexts underscores its evolutionary significance while revealing novel targets for resistance management, particularly biotin-based synergists and AhR dimerization inhibitors.
杀虫剂抗药性对全球农业可持续性构成重大挑战。虽然代谢解毒和靶点突变是抗性机制的特征,但微量营养素稳态的作用仍未得到充分研究。本研究揭示了虫螺生物素缺乏通过协调化学感觉蛋白(CSP)动态的多层调控网络驱动吡虫啉抗性。生物素缺乏通过与抗性菌株中过表达的CSP2、CSP4、CSP7和CSP15的高亲和力结合,增强了csp介导的杀虫剂隔离。RNA干扰和双荧光素酶实验表明,芳烃受体及其核转运子(AhR/ARNT)转录激活CSP2和CSP15,其敲除部分恢复了对杀虫剂的敏感性。此外,生物素缺乏激活活性氧(ROS)/cap 'n' collar C (CncC)信号,通过转录重编程提高AhR/ARNT的表达。酵母三杂交实验鉴定了翻译后调控层,其中生物素直接抑制AhR-ARNT异源二聚化。田间进化的抗性群体概括了这一机制,表现出保守的分子特征,包括生物素缺乏、ROS/CncC途径激活以及与抗性强度相关的AhR/ARNT-CSP过表达。这些发现建立了一个统一的模型,其中生物素稀缺性通过三种协同机制重新编程外源防御:增强csp -杀虫剂结合,通过ROS/ cc - ahr /ARNT信号进行转录扩增,转录复合物的翻译后优化。该途径在实验室和生态环境中的操作保护强调了其进化意义,同时揭示了耐药性管理的新靶点,特别是基于生物素的增效剂和AhR二聚化抑制剂。
{"title":"Biotin deficiency bridges metabolic signaling to insecticide sequestration in Nilaparvata lugens","authors":"Mengqing Deng, Xiyue Xu, Zhiming Yang, Kai Lu","doi":"10.1016/j.ibmb.2025.104445","DOIUrl":"10.1016/j.ibmb.2025.104445","url":null,"abstract":"<div><div>Insecticide resistance poses a critical challenge to global agricultural sustainability. While metabolic detoxification and target-site mutations are well-characterized resistance mechanisms, the role of micronutrient homeostasis remains understudied. This study reveals that biotin deficiency in <em>Nilaparvata lugens</em> drives imidacloprid resistance through a multi-tiered regulatory network coordinating chemosensory protein (CSP) dynamics. Biotin deficiency enhances CSP-mediated insecticide sequestration via high-affinity binding to CSP2, CSP4, CSP7, and CSP15, which are overexpressed in resistant strains. RNA interference and dual-luciferase assays demonstrate that the aryl hydrocarbon receptor and its nuclear translocator (AhR/ARNT) transcriptionally activate <em>CSP2</em> and <em>CSP15</em>, with their knockdown partially restoring insecticide susceptibility. Furthermore, biotin deficiency activates reactive oxygen species (ROS)/cap ‘n’ collar C (CncC) signaling, elevating <em>AhR/ARNT</em> expression through transcriptional reprogramming. Yeast three-hybrid assays identify a post-translational regulatory layer, wherein biotin directly inhibits AhR–ARNT heterodimerization. Field-evolved resistant populations recapitulate this mechanism, exhibiting conserved molecular signatures including biotin deficiency, ROS/CncC pathway activation, and AhR/ARNT-CSP overexpression correlated with resistance intensity. These findings establish a unified model wherein biotin scarcity reprograms xenobiotic defense through three synergistic mechanisms: enhanced CSP–insecticide binding, transcriptional amplification via ROS/CncC-AhR/ARNT signaling, and post-translational optimization of transcriptional complexes. The operational conservation of this pathway across laboratory and ecological contexts underscores its evolutionary significance while revealing novel targets for resistance management, particularly biotin-based synergists and AhR dimerization inhibitors.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104445"},"PeriodicalIF":3.7,"publicationDate":"2025-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145487357","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 : 2025-11-07DOI: 10.1016/j.ibmb.2025.104442
Lifang Li , Jingyi Ning , Tao Zhu , Mingxian Lan , Mehboob Hussain , Xi Gao , Jiaying Zhu , Guoxing Wu
The tephritid stem gall fly, Procecidochares utilis Stone (Diptera: Trypetidae), serves as an effective biocontrol agent against the invasive plant Eupatorium adenophorum (syn. Ageratina adenophorum) by inducing galls through larval saliva secretions, thereby inhibiting the growth of E. adenophorum. Despite this, the genetic basis of gall induction by P. utilis remains unexplored. This study generated a chromosome-level genome of P. utilis, totaling 467.64 Mb with a scaffold N50 of 74.29 Mb, organized into six pseudochromosomes. Comparative genomic analysis revealed a significant expansion of apolipoprotein D (ApoD) genes—a novel finding among gall-forming insects. Salivary gland transcriptomics identified 124 putative secreted proteins, with ApoD expansions exhibiting predominant expression. Among 29 identified ApoD genes in the P. utilis genome, 24 formed a species-specific clade, predominantly expressed in larvae and salivary glands. Notably, 21 of these genes were associated with salivary gland secretion. PuApoD311, uniquely expressed in P. utilis larvae and salivary glands, was successfully recombinantly produced in Sf9 cells using the baculovirus system. Injection of recombinant PuApoD311 into E. adenophorum resulted in the formation of “gall-like structures, characterized by analogous lateral root proliferation” structures, and elevated zeatin levels, indicating its pivotal role in gall formation. These findings contribute to our comprehension of salivary gland protein evolution in P. utilis and the molecular mechanisms governing gall induction by this biocontrol agent.
{"title":"Involvement of salivary Apolipoprotein D, PuApoD311, in gall formation induced by the tephritid stem gall fly, Procecidochares utilis","authors":"Lifang Li , Jingyi Ning , Tao Zhu , Mingxian Lan , Mehboob Hussain , Xi Gao , Jiaying Zhu , Guoxing Wu","doi":"10.1016/j.ibmb.2025.104442","DOIUrl":"10.1016/j.ibmb.2025.104442","url":null,"abstract":"<div><div>The tephritid stem gall fly, <em>Procecidochares utilis</em> Stone (Diptera: Trypetidae), serves as an effective biocontrol agent against the invasive plant <em>Eupatorium adenophorum</em> (syn. <em>Ageratina adenophorum</em>) by inducing galls through larval saliva secretions, thereby inhibiting the growth of <em>E. adenophorum</em>. Despite this, the genetic basis of gall induction by <em>P. utilis</em> remains unexplored. This study generated a chromosome-level genome of <em>P. utilis</em>, totaling 467.64 Mb with a scaffold N50 of 74.29 Mb, organized into six pseudochromosomes. Comparative genomic analysis revealed a significant expansion of <em>apolipoprotein D</em> (<em>ApoD</em>) genes—a novel finding among gall-forming insects. Salivary gland transcriptomics identified 124 putative secreted proteins, with <em>ApoD</em> expansions exhibiting predominant expression. Among 29 identified <em>ApoD</em> genes in the <em>P. utilis</em> genome, 24 formed a species-specific clade, predominantly expressed in larvae and salivary glands. Notably, 21 of these genes were associated with salivary gland secretion. <em>PuApoD311</em>, uniquely expressed in <em>P. utilis</em> larvae and salivary glands, was successfully recombinantly produced in Sf9 cells using the baculovirus system. Injection of recombinant <em>PuApoD311</em> into <em>E. adenophorum</em> resulted in the formation of “gall-like structures, characterized by analogous lateral root proliferation” structures, and elevated zeatin levels, indicating its pivotal role in gall formation. These findings contribute to our comprehension of salivary gland protein evolution in <em>P. utilis</em> and the molecular mechanisms governing gall induction by this biocontrol agent.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104442"},"PeriodicalIF":3.7,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145480491","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 : 2025-11-07DOI: 10.1016/j.ibmb.2025.104443
Jin Myung Kim , Bo Yeon Kim , Yun Hui Kim , Hyung Joo Yoon , Yong Soo Choi , Kyeong Yong Lee , Dong Won Kim , Kwang Sik Lee , Byung Rae Jin
The Niemann–Pick type C2 protein (NPC2) functions as a cholesterol modulator in the sperm membrane and enhances sperm physiological functions in vertebrates and some invertebrates. In insects, NPC2 is involved in lipid metabolism, immune response signaling, and chemical communication. However, the role of NPC2 as a sperm-associated protein in insect sperm remains to be elucidated. This study identified a novel function of the Apis mellifera NPC2 protein (AmNPC2) as a sperm-binding protein. Immunoassays and binding assays using recombinant AmNPC2 protein and its specific antibody revealed that AmNPC2 is expressed in the testes of drones and is localized on the sperm surface as a sperm-binding protein. Furthermore, a reduction in AmNPC2 levels on the sperm surface decreased sperm viability. AmNPC2 also appeared to play a protective role in maintaining sperm viability under both oxidative and temperature stress conditions. Our findings indicate that AmNPC2 is a sperm-binding protein that enhances the viability of honeybee sperm.
{"title":"The role of the Niemann–Pick type C2 protein as a sperm-binding protein in honeybees","authors":"Jin Myung Kim , Bo Yeon Kim , Yun Hui Kim , Hyung Joo Yoon , Yong Soo Choi , Kyeong Yong Lee , Dong Won Kim , Kwang Sik Lee , Byung Rae Jin","doi":"10.1016/j.ibmb.2025.104443","DOIUrl":"10.1016/j.ibmb.2025.104443","url":null,"abstract":"<div><div>The Niemann–Pick type C2 protein (NPC2) functions as a cholesterol modulator in the sperm membrane and enhances sperm physiological functions in vertebrates and some invertebrates. In insects, NPC2 is involved in lipid metabolism, immune response signaling, and chemical communication. However, the role of NPC2 as a sperm-associated protein in insect sperm remains to be elucidated. This study identified a novel function of the <em>Apis mellifera</em> NPC2 protein (AmNPC2) as a sperm-binding protein. Immunoassays and binding assays using recombinant AmNPC2 protein and its specific antibody revealed that AmNPC2 is expressed in the testes of drones and is localized on the sperm surface as a sperm-binding protein. Furthermore, a reduction in AmNPC2 levels on the sperm surface decreased sperm viability. AmNPC2 also appeared to play a protective role in maintaining sperm viability under both oxidative and temperature stress conditions. Our findings indicate that AmNPC2 is a sperm-binding protein that enhances the viability of honeybee sperm.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104443"},"PeriodicalIF":3.7,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145480450","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 : 2025-11-06DOI: 10.1016/j.ibmb.2025.104440
Huamei Wen , Xinyue Ma , Wenjing Zhang , Lu Wang , Yanbin Li , Dan Lv , Jianghua Sun , Dingze Mang
Fructose is an important energy source for many insects, yet its perception and physiological roles in the red imported fire ant, Solenopsis invicta, remain unclear. Here, we show that S. invicta workers strongly prefer sucrose and glucose but demonstrate only weak attraction for fructose. Using Ca2+ imaging and intracellular Ca2+ assays, we identified SinvGr43a as a fructose-responsive gustatory receptor. While its expression is low in the antennae, reverse transcription-quantitative PCR (RT-qPCR) revealed high levels in the brain and gut, suggesting an internal sensing role. RNA interference-mediated knockdown of SinvGr43a led to reduced production and secretion of short neuropeptide F (sNPF) in the gut and insulin-like peptide (ILP) in both the brain and gut. Moreover, this silencing also downregulated key fatty acid metabolic regulators, including sterol regulatory element-binding protein (SREBP) and fatty acid synthase (FAS), in the fat body, resulting in decreased lipid storage. Our findings demonstrate that SinvGr43a functions primarily as an internal fructose sensor, linking nutrient detection with neuropeptide signaling and lipid metabolism to maintain energy homeostasis in S. invicta.
{"title":"A fructose-sensitive gustatory receptor links nutrient sensing to lipid metabolism in the red imported fire ant","authors":"Huamei Wen , Xinyue Ma , Wenjing Zhang , Lu Wang , Yanbin Li , Dan Lv , Jianghua Sun , Dingze Mang","doi":"10.1016/j.ibmb.2025.104440","DOIUrl":"10.1016/j.ibmb.2025.104440","url":null,"abstract":"<div><div>Fructose is an important energy source for many insects, yet its perception and physiological roles in the red imported fire ant, <em>Solenopsis invicta</em>, remain unclear. Here, we show that <em>S. invicta</em> workers strongly prefer sucrose and glucose but demonstrate only weak attraction for fructose. Using Ca<sup>2+</sup> imaging and intracellular Ca<sup>2+</sup> assays, we identified <em>SinvGr43a</em> as a fructose-responsive gustatory receptor. While its expression is low in the antennae, reverse transcription-quantitative PCR (RT-qPCR) revealed high levels in the brain and gut, suggesting an internal sensing role. RNA interference-mediated knockdown of <em>SinvGr43a</em> led to reduced production and secretion of short neuropeptide F (sNPF) in the gut and insulin-like peptide (ILP) in both the brain and gut. Moreover, this silencing also downregulated key fatty acid metabolic regulators, including sterol regulatory element-binding protein (SREBP) and fatty acid synthase (FAS), in the fat body, resulting in decreased lipid storage. Our findings demonstrate that <em>SinvGr43a</em> functions primarily as an internal fructose sensor, linking nutrient detection with neuropeptide signaling and lipid metabolism to maintain energy homeostasis in <em>S. invicta</em>.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104440"},"PeriodicalIF":3.7,"publicationDate":"2025-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145476501","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 : 2025-11-06DOI: 10.1016/j.ibmb.2025.104441
Maithili Datta , Katrin Luck , Song Wu , Feng Chen , Yuko E. Ulrich , Sarah E. O'Connor , Tobias G. Köllner
Iridoids are bicyclic monoterpenoids that function as defense and signaling compounds in both plants and insects. Although recent research suggested that iridoids evolved independently in these two kingdoms of life, it remained unclear whether independent evolution of iridoid biosynthesis also occurred across different insect lineages. In order to gain insight into the evolution of iridoids in insects, we examined the first committed step of iridoid biosynthesis, the hydroxylation of geraniol to 8-hydroxygeraniol, in the Argentine ant (Linepithema humile) of the order Hymenoptera. This transformation is typically catalyzed by cytochrome P450 monooxygenases in both plants and insects. By integrating transcriptomic and metabolomic analyses across various body parts, we identified candidate P450 genes potentially involved in this reaction. The candidate genes were heterologously expressed in yeast, and enzyme activity was assessed by supplying geraniol to the yeast cultures. One candidate P450 enzyme displayed geraniol 8-hydroxylase (G8H) activity and was designated LhG8H. Phylogenetic analysis showed that LhG8H is evolutionarily distinct from previously characterized G8H enzymes in the pea aphid (Acyrthosiphon pisum) of the order Hemiptera and the mustard leaf beetle (Phaedon cochleariae) of the order Coleoptera. These results support the hypothesis that geraniol 8-hydroxylase activity, a key step in iridoid biosynthesis, evolved independently within the insect orders Hymenoptera, Hemiptera, and Coleoptera.
{"title":"Independent evolution of geraniol-8-hydroxylase activity involved in iridoid formation in the Argentine ant (Linepithema humile)","authors":"Maithili Datta , Katrin Luck , Song Wu , Feng Chen , Yuko E. Ulrich , Sarah E. O'Connor , Tobias G. Köllner","doi":"10.1016/j.ibmb.2025.104441","DOIUrl":"10.1016/j.ibmb.2025.104441","url":null,"abstract":"<div><div>Iridoids are bicyclic monoterpenoids that function as defense and signaling compounds in both plants and insects. Although recent research suggested that iridoids evolved independently in these two kingdoms of life, it remained unclear whether independent evolution of iridoid biosynthesis also occurred across different insect lineages. In order to gain insight into the evolution of iridoids in insects, we examined the first committed step of iridoid biosynthesis, the hydroxylation of geraniol to 8-hydroxygeraniol, in the Argentine ant (<em>Linepithema humile</em>) of the order Hymenoptera. This transformation is typically catalyzed by cytochrome P450 monooxygenases in both plants and insects. By integrating transcriptomic and metabolomic analyses across various body parts, we identified candidate P450 genes potentially involved in this reaction. The candidate genes were heterologously expressed in yeast, and enzyme activity was assessed by supplying geraniol to the yeast cultures. One candidate P450 enzyme displayed geraniol 8-hydroxylase (G8H) activity and was designated LhG8H. Phylogenetic analysis showed that LhG8H is evolutionarily distinct from previously characterized G8H enzymes in the pea aphid (<em>Acyrthosiphon pisum</em>) of the order Hemiptera and the mustard leaf beetle (<em>Phaedon cochleariae</em>) of the order Coleoptera. These results support the hypothesis that geraniol 8-hydroxylase activity, a key step in iridoid biosynthesis, evolved independently within the insect orders Hymenoptera, Hemiptera, and Coleoptera.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104441"},"PeriodicalIF":3.7,"publicationDate":"2025-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145475253","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 : 2025-11-04DOI: 10.1016/j.ibmb.2025.104433
Rong Hu , Jun Wu , Siying Li , Peiyu Yang , Gang Wu , Changying Niu , Shuai Zhan , Yazhou Chen
Horizontal gene transfer (HGT) plays a crucial role in genome evolution, especially when it enables the acquisition and assembly of multi-step biosynthetic pathways. Here, we investigate the evolutionary origins of carotenoid biosynthesis genes in aphids to determine whether multiple functionally related genes were acquired through HGT. We analyzed carotenoid biosynthesis genes in 23 aphid genomes based on homologs in plants, fungi, and bacteria. Phylogenetic analyses revealed that Geranylgeranyl pyrophosphate synthase (GPS), Phytoene synthase (PS), and Carotenoid desaturase (CD) were acquired via HGT from fungi by ancestral insect species, while Carotenoid cleavage oxygenase (CCO) appears to be a native insect gene. Most insect genomes contain two GPS copies, likely resulting from independent HGT events, whereas aphid genomes exhibit extensive duplication of PS and CD, a pattern uncommon in other insects. Expression analyses across aphid species with distinct pigmentation showed that these genes are broadly transcribed with substantial variability in expression levels. In Myzus persicae, comparative expression analysis between red and green clones, as well as a green-red clone with green and red color polymorphism, revealed that PS-4390 is a novel candidate for red pigmentation in M. persicae, in addition to CD-4400, a homolog of the tor gene in Acyrthosiphon pisum. These findings provide strong evidence that HGT can introduce multiple functionally related genes into recipient genomes, allowing them to be co-opted into a functional biosynthetic pathway.
{"title":"Coordinated horizontal transfer of multiple genes assembles a carotenoid biosynthesis pathway in aphids","authors":"Rong Hu , Jun Wu , Siying Li , Peiyu Yang , Gang Wu , Changying Niu , Shuai Zhan , Yazhou Chen","doi":"10.1016/j.ibmb.2025.104433","DOIUrl":"10.1016/j.ibmb.2025.104433","url":null,"abstract":"<div><div>Horizontal gene transfer (HGT) plays a crucial role in genome evolution, especially when it enables the acquisition and assembly of multi-step biosynthetic pathways. Here, we investigate the evolutionary origins of carotenoid biosynthesis genes in aphids to determine whether multiple functionally related genes were acquired through HGT. We analyzed carotenoid biosynthesis genes in 23 aphid genomes based on homologs in plants, fungi, and bacteria. Phylogenetic analyses revealed that <em>Geranylgeranyl pyrophosphate synthase</em> (<em>GPS</em>), <em>Phytoene synthase</em> (<em>PS</em>), and <em>Carotenoid desaturase</em> (<em>CD</em>) were acquired via HGT from fungi by ancestral insect species, while <em>Carotenoid cleavage oxygenase</em> (<em>CCO</em>) appears to be a native insect gene. Most insect genomes contain two <em>GPS</em> copies, likely resulting from independent HGT events, whereas aphid genomes exhibit extensive duplication of <em>PS</em> and <em>CD</em>, a pattern uncommon in other insects. Expression analyses across aphid species with distinct pigmentation showed that these genes are broadly transcribed with substantial variability in expression levels. In <em>Myzus persicae</em>, comparative expression analysis between red and green clones, as well as a green-red clone with green and red color polymorphism, revealed that <em>PS-4390</em> is a novel candidate for red pigmentation in <em>M. persicae</em>, in addition to <em>CD-4400</em>, a homolog of the <em>tor</em> gene in <em>Acyrthosiphon pisum</em>. These findings provide strong evidence that HGT can introduce multiple functionally related genes into recipient genomes, allowing them to be co-opted into a functional biosynthetic pathway.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"186 ","pages":"Article 104433"},"PeriodicalIF":3.7,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145457032","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}