Gain-of-function analysis is critical to directly assessing gene functions but remains challenging in non-traditional model insects. Recent development of the hyperactive piggyBac transposase (hyPBase) has improved the efficiency of germline transformation in ametabolous and holometabolous insects. Here, we report highly efficient somatic and germline transformations mediated by hyPBase in the cricket Gryllus bimaculatus. hyPBase-mediated misexpression of the melanin pathway gene arylalkylamine N-Acetyltransferase (aaNAT) turned the cricket body color from black to a light color, indicating a broad effect on external phenotype in the injected generation. Mosaic expression of the homeotic gene Antennapedia with hyPBase successfully circumvented the potential embryonic lethality, and induced antenna-to-leg homeotic transformation in a hatched nymph. Our data provide a practical methodology for gain-of-function analysis in the hemimetabolous model to understand key evolutionary innovations in insects.
{"title":"Highly efficient hyPBase-mediated transgenesis facilitates gain-of-function analysis in the cricket Gryllus bimaculatus","authors":"Takahiro Ohde , Taro Nakamura , Taro Mito , Takaaki Daimon","doi":"10.1016/j.ibmb.2025.104405","DOIUrl":"10.1016/j.ibmb.2025.104405","url":null,"abstract":"<div><div>Gain-of-function analysis is critical to directly assessing gene functions but remains challenging in non-traditional model insects. Recent development of the hyperactive <em>piggyBac</em> transposase (hyPBase) has improved the efficiency of germline transformation in ametabolous and holometabolous insects. Here, we report highly efficient somatic and germline transformations mediated by hyPBase in the cricket <em>Gryllus bimaculatus</em>. hyPBase-mediated misexpression of the melanin pathway gene <em>arylalkylamine N-Acetyltransferase</em> (<em>aaNAT</em>) turned the cricket body color from black to a light color, indicating a broad effect on external phenotype in the injected generation. Mosaic expression of the homeotic gene <em>Antennapedia</em> with hyPBase successfully circumvented the potential embryonic lethality, and induced antenna-to-leg homeotic transformation in a hatched nymph. Our data provide a practical methodology for gain-of-function analysis in the hemimetabolous model to understand key evolutionary innovations in insects.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104405"},"PeriodicalIF":3.7,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145155184","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 : 2025-09-22DOI: 10.1016/j.ibmb.2025.104402
Huitang Qi, Tian Liu
The insect cuticle is essential for survival and environmental adaptation, comprising over 100 cuticular proteins (CPs) that represent approximately 1 % of the insect genome coding genes. This complexity is unique among biological materials, underscoring the diverse roles of CPs in insect biology. However, the reasons for this extensive CP repertoire and their precise functional mechanisms persist as unresolved questions in entomology. Recently, advances in molecular biology and genomics have yielded significant progress in elucidating CP functions. Studies demonstrate that CPs are crucial not only for cuticle formation and development but also for regulating body size, coloration, cuticular structure development, environmental adaptation, immune defense, viral interactions, and insecticide resistance. This review comprehensively summarizes current insect CP research, highlights their diverse physiological and ecological roles, and explores their translational potential for pest management and biomimetic material design.
{"title":"Cuticular proteins: Essential molecular code for insect survival","authors":"Huitang Qi, Tian Liu","doi":"10.1016/j.ibmb.2025.104402","DOIUrl":"10.1016/j.ibmb.2025.104402","url":null,"abstract":"<div><div>The insect cuticle is essential for survival and environmental adaptation, comprising over 100 cuticular proteins (CPs) that represent approximately 1 % of the insect genome coding genes. This complexity is unique among biological materials, underscoring the diverse roles of CPs in insect biology. However, the reasons for this extensive CP repertoire and their precise functional mechanisms persist as unresolved questions in entomology. Recently, advances in molecular biology and genomics have yielded significant progress in elucidating CP functions. Studies demonstrate that CPs are crucial not only for cuticle formation and development but also for regulating body size, coloration, cuticular structure development, environmental adaptation, immune defense, viral interactions, and insecticide resistance. This review comprehensively summarizes current insect CP research, highlights their diverse physiological and ecological roles, and explores their translational potential for pest management and biomimetic material design.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104402"},"PeriodicalIF":3.7,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145118624","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 : 2025-09-22DOI: 10.1016/j.ibmb.2025.104403
Benjamin J. Hunt , Kumar Saurabh Singh , Bartlomiej J. Troczka , Joanna Mackisack , Emma Randall , Mark Mallott , Tobias Baril , James Galbraith , Bram Kuijper , Ralf Nauen , Alex Hayward , Chris Bass
Insect pests evolve heritable resistance to insecticides through selection acting on genetic variation generated by mutation. It has been hypothesised that exposure to low doses of pesticides may increase mutation rate in pest genomes and accelerate resistance evolution. However, the impact of pesticide-induced stress on the mutation rate of insects has never been empirically tested. Here we leverage new, high-quality genomic resources for the aphid pest Myzus persicae in conjunction with long-term mutation accumulation experiments to interrogate spontaneous genetic and epigenetic mutation rates in insecticide-exposed and unexposed aphid lines. Our data reveal that multigenerational exposure of Myzus persicae to sublethal concentrations of the neonicotinoid insecticide imidacloprid does not increase genetic mutation rate. Rather we show that imidacloprid exposure results in a significantly lower epimutation rate. These findings reveal the rate of origin and composition of (epi)mutations arising in a pest insect and challenge the proposed link between pesticide exposure and the rate of mutation.
{"title":"Sub-lethal insecticide stress alters epimutation rate but not genetic mutation rate in the pest insect Myzus persicae","authors":"Benjamin J. Hunt , Kumar Saurabh Singh , Bartlomiej J. Troczka , Joanna Mackisack , Emma Randall , Mark Mallott , Tobias Baril , James Galbraith , Bram Kuijper , Ralf Nauen , Alex Hayward , Chris Bass","doi":"10.1016/j.ibmb.2025.104403","DOIUrl":"10.1016/j.ibmb.2025.104403","url":null,"abstract":"<div><div>Insect pests evolve heritable resistance to insecticides through selection acting on genetic variation generated by mutation. It has been hypothesised that exposure to low doses of pesticides may increase mutation rate in pest genomes and accelerate resistance evolution. However, the impact of pesticide-induced stress on the mutation rate of insects has never been empirically tested. Here we leverage new, high-quality genomic resources for the aphid pest <em>Myzus persicae</em> in conjunction with long-term mutation accumulation experiments to interrogate spontaneous genetic and epigenetic mutation rates in insecticide-exposed and unexposed aphid lines. Our data reveal that multigenerational exposure of <em>Myzus persicae</em> to sublethal concentrations of the neonicotinoid insecticide imidacloprid does not increase genetic mutation rate. Rather we show that imidacloprid exposure results in a significantly lower epimutation rate. These findings reveal the rate of origin and composition of (epi)mutations arising in a pest insect and challenge the proposed link between pesticide exposure and the rate of mutation.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104403"},"PeriodicalIF":3.7,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145135902","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 : 2025-09-17DOI: 10.1016/j.ibmb.2025.104401
Tong-Jun Jin , Zheng-Yang Wang , Qian-Ping Xie , Li-Yuan Zheng , Guy Smagghe , Jin-Jun Wang , Qiang Zhang , Wei Dou
The interplay between lipid homeostasis and hormonal regulation is critical for reproductive success in female insects. The molecular mechanisms linking lipid metabolism to hormonal pathways, particularly ecdysteroid synthesis, remain poorly understood. In this study, we reveal a novel regulatory role of miR-927 in the oriental fruit fly, Bactrocera dorsalis. Using in vivo miRNA mimic/inhibitor assays, combined with an in vitro dual-luciferase reporter and RNA pull-down experiments, we demonstrated that miR-927 directly targets BdCYP315a1, a key enzyme in ecdysteroidogenesis, thereby repressing the 20-hydroxyecdysone (20E) levels. RNA interference of BdCYP315a1 confirmed its essential role in maintaining 20E homeostasis and proper developmental progression. Further, we show that miR-927 expression is regulated by the insulin/insulin-like signaling (IIS) pathway, linking nutritional status to hormonal control. Changes in miR-927, BdCYP315a1, and 20E levels corresponded with alterations in lipid content, ultimately influencing fecundity. These findings reveal a previously uncharacterized miRNA-mediated network integrating nutritional signals, hormonal regulation, and lipid metabolism to control reproductive outcomes. Our work provides novel insights into the molecular mechanisms underlying insect reproduction, highlights miRNAs as key mediators of hormone-lipid interactions, and offers valuable resources for identifying miRNA targets in non-model insect species.
{"title":"miR-927 links nutrient signals and 20-hydroxyecdysone regulation and mediates oviposition in Bactrocera dorsalis","authors":"Tong-Jun Jin , Zheng-Yang Wang , Qian-Ping Xie , Li-Yuan Zheng , Guy Smagghe , Jin-Jun Wang , Qiang Zhang , Wei Dou","doi":"10.1016/j.ibmb.2025.104401","DOIUrl":"10.1016/j.ibmb.2025.104401","url":null,"abstract":"<div><div>The interplay between lipid homeostasis and hormonal regulation is critical for reproductive success in female insects. The molecular mechanisms linking lipid metabolism to hormonal pathways, particularly ecdysteroid synthesis, remain poorly understood. In this study, we reveal a novel regulatory role of miR-927 in the oriental fruit fly, <em>Bactrocera dorsalis</em>. <em>U</em>sing <em>in vivo</em> miRNA mimic/inhibitor assays, combined with an <em>in vitro</em> dual-luciferase reporter and RNA pull-down experiments, we demonstrated that miR-927 directly targets <em>BdCYP315a1</em>, a key enzyme in ecdysteroidogenesis, thereby repressing the 20-hydroxyecdysone (20E) levels. RNA interference of <em>BdCYP315a1</em> confirmed its essential role in maintaining 20E homeostasis and proper developmental progression. Further, we show that miR-927 expression is regulated by the insulin/insulin-like signaling (IIS) pathway, linking nutritional status to hormonal control. Changes in miR-927, <em>BdCYP315a1</em>, and 20E levels corresponded with alterations in lipid content, ultimately influencing fecundity. These findings reveal a previously uncharacterized miRNA-mediated network integrating nutritional signals, hormonal regulation, and lipid metabolism to control reproductive outcomes. Our work provides novel insights into the molecular mechanisms underlying insect reproduction, highlights miRNAs as key mediators of hormone-lipid interactions, and offers valuable resources for identifying miRNA targets in non-model insect species.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104401"},"PeriodicalIF":3.7,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145090911","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 : 2025-09-10DOI: 10.1016/j.ibmb.2025.104400
Dries Amezian, Thomas Van Leeuwen
The CncC/Keap1 signalling pathway regulates antioxidant and detoxification gene expression in arthropods and is frequently associated with metabolic insecticide resistance. This review critically assesses evidence for its role in resistance phenotypes across key pest species. Although overactivation of CncC/Keap1 correlates with increased detoxification enzyme expression and pesticide tolerance, causal mutations in the coding or regulatory regions of CncC, Keap1, or Maf remain unidentified. We evaluate the evidence supporting the role of CncC/Keap1 in pesticide resistance in insects and mites and report the latest advancements in our understanding of this system in arthropods. We further highlight the need for unbiased genetic mapping and reverse genetic approaches to resolve the mechanisms of constitutive CncC activation in resistant populations. Understanding these mechanisms is crucial for elucidating the origins of metabolic resistance and developing sustainable pest management strategies.
{"title":"Is the CncC/Keap1 complex a major factor in conferring pesticide resistance in arthropods? - A critical review","authors":"Dries Amezian, Thomas Van Leeuwen","doi":"10.1016/j.ibmb.2025.104400","DOIUrl":"10.1016/j.ibmb.2025.104400","url":null,"abstract":"<div><div>The CncC/Keap1 signalling pathway regulates antioxidant and detoxification gene expression in arthropods and is frequently associated with metabolic insecticide resistance. This review critically assesses evidence for its role in resistance phenotypes across key pest species. Although overactivation of CncC/Keap1 correlates with increased detoxification enzyme expression and pesticide tolerance, causal mutations in the coding or regulatory regions of <em>CncC</em>, <em>Keap1</em>, or <em>Maf</em> remain unidentified. We evaluate the evidence supporting the role of CncC/Keap1 in pesticide resistance in insects and mites and report the latest advancements in our understanding of this system in arthropods. We further highlight the need for unbiased genetic mapping and reverse genetic approaches to resolve the mechanisms of constitutive CncC activation in resistant populations. Understanding these mechanisms is crucial for elucidating the origins of metabolic resistance and developing sustainable pest management strategies.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104400"},"PeriodicalIF":3.7,"publicationDate":"2025-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145045149","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 : 2025-09-09DOI: 10.1016/j.ibmb.2025.104399
Hanwen Ye , Chanqin Zheng , Bing Wang , Yuqing Zhu , Chenjing Wang , Zhuobing Wang , Zhanjun Lu , Weiyi He , Wei Chen
The diamondback moth (Plutella xylostella), a globally destructive pest, has Brassicaceae as its long-term co-evolved host and can also utilize Fabaceae as an alternative field host. The primary differential factor between these plant families is glucosinolates (GLs). Conventional transcriptome data revealed high midgut expression of glucosinolate sulfatases (GSSs) in response to glucosinolates. However, due to the inability of conventional transcriptomics to resolve cellular heterogeneity, it remained unclear which specific cell types express GSSs and whether exposure to different field hosts triggers distinct differentiation fates in these cells. To address these questions, we constructed a high-resolution atlas of 28,451 midgut cells from larvae reared on radish and pea leaves. Marker gene-based clustering identified 16 distinct cell types, including enterocytes (ECs), enteroendocrine cells (EEs), and intestinal stem cells (ISCs). ScRNA-seq and qRT-PCR analyses revealed host-dependent differential expression patterns of three GSS genes (GSS1-GSS3) among ECs, EEs, and ISCs. Notably, elevated GSS3 protein levels in radish-fed larvae were confirmed by Western blot (WB) and Immunohistochemistry (IHC), while GSS1 and GSS2 exhibited more variable expression patterns. Pseudo-time trajectory analysis further demonstrated that all three GSSs followed differentiation pathways from ISCs into EEs or ECs, but dietary conditions led to distinct differentiation trajectories. This study elucidates the diet-dependent regulatory landscape of GSSs in P. xylostella and delineates the differentiation trajectory of GSS-expressing cells.
小菜蛾（Plutella xylostella）是一种全球性的破坏性害虫，它以十字花科作为长期共同进化的寄主，也可以利用豆科作为替代的田间寄主。这些植物家族之间的主要差异因素是硫代葡萄糖苷（GLs）。常规转录组数据显示，在对硫代葡萄糖苷的反应中，中肠高表达硫代葡萄糖苷硫酸酯酶（gss）。然而，由于传统转录组学无法解决细胞异质性，目前尚不清楚哪些特定细胞类型表达gss，以及暴露于不同的野宿主是否会触发这些细胞的不同分化命运。为了解决这些问题，我们构建了28,451个中肠细胞的高分辨率图谱，这些细胞来自萝卜和豌豆叶上饲养的幼虫。基于标记基因的聚类鉴定出16种不同的细胞类型，包括肠细胞（ECs）、肠内分泌细胞（EEs）和肠干细胞（ISCs）。ScRNA-seq和qRT-PCR分析揭示了三个GSS基因（GSS1-GSS3）在ECs、EEs和ISCs中宿主依赖性的差异表达模式。Western blot （WB）和免疫组化（IHC）结果显示，萝卜饲幼虫中GSS3蛋白水平升高，而GSS1和GSS2表达模式变化较大。伪时间轨迹分析进一步表明，所有三种gss都遵循从ISCs到EEs或ECs的分化途径，但饮食条件导致不同的分化轨迹。本研究阐明了小菜中gss的饮食依赖性调控格局，并描绘了gss表达细胞的分化轨迹。
{"title":"Single-cell transcriptomic profiling reveals diet-dependent dynamics of glucosinolate sulfatases expression and cellular origin in the midgut of Plutella xylostella","authors":"Hanwen Ye , Chanqin Zheng , Bing Wang , Yuqing Zhu , Chenjing Wang , Zhuobing Wang , Zhanjun Lu , Weiyi He , Wei Chen","doi":"10.1016/j.ibmb.2025.104399","DOIUrl":"10.1016/j.ibmb.2025.104399","url":null,"abstract":"<div><div>The diamondback moth (<em>Plutella xylostella</em>), a globally destructive pest, has Brassicaceae as its long-term co-evolved host and can also utilize Fabaceae as an alternative field host. The primary differential factor between these plant families is glucosinolates (GLs). Conventional transcriptome data revealed high midgut expression of <em>glucosinolate sulfatases</em> (<em>GSSs</em>) in response to glucosinolates. However, due to the inability of conventional transcriptomics to resolve cellular heterogeneity, it remained unclear which specific cell types express <em>GSSs</em> and whether exposure to different field hosts triggers distinct differentiation fates in these cells. To address these questions, we constructed a high-resolution atlas of 28,451 midgut cells from larvae reared on radish and pea leaves. Marker gene-based clustering identified 16 distinct cell types, including enterocytes (ECs), enteroendocrine cells (EEs), and intestinal stem cells (ISCs). ScRNA-seq and qRT-PCR analyses revealed host-dependent differential expression patterns of three <em>GSS</em> genes (<em>GSS1-GSS3</em>) among ECs, EEs, and ISCs. Notably, elevated GSS3 protein levels in radish-fed larvae were confirmed by Western blot (WB) and Immunohistochemistry (IHC), while GSS1 and GSS2 exhibited more variable expression patterns. Pseudo-time trajectory analysis further demonstrated that all three <em>GSSs</em> followed differentiation pathways from ISCs into EEs or ECs, but dietary conditions led to distinct differentiation trajectories. This study elucidates the diet-dependent regulatory landscape of <em>GSSs</em> in <em>P. xylostella</em> and delineates the differentiation trajectory of <em>GSS</em>-expressing cells.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104399"},"PeriodicalIF":3.7,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145032506","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 : 2025-09-08DOI: 10.1016/j.ibmb.2025.104397
Kun-Yu Yang, Yi-Fei Sun, Yuan-Sen Liang, Hao Li, Meng-Xue Qi, Zhaowei Wang, Luis Carlos Ramos Aguila, Li-Qun Cai, Hao-Sen Li, Hong Pang
Horizontal gene transfer (HGT) is now widely recognized as an important mechanism contributing to host immunity and adaptation. Ladybird beetles, with their diverse diets and habitats, encounter a broad spectrum of microbial threats, making effective immune responses critical for their survival. However, the immune roles of HGT-acquired genes in ladybirds remain largely unexplored. To address this gap, we investigated HGT of a NADAR (NAD- and ADP-ribose-associated) domain-containing gene from microorganisms to insects. Phylogenetic analyses revealed that NADAR genes in ladybird beetles form a well-supported clade nested within a larger group composed primarily of bacterial sequences, providing strong evidence for an HGT origin. Sampling across 69 ladybird species suggests that NADAR genes originated in the Coccinellidae family and were subsequently retained or duplicated across ladybird genomes, indicating their functional importance. Using the ladybird Cryptolaemus montrouzieri as a model, we observed that the expression levels of CmNADAR1 and CmNADAR2 were significantly upregulated in response to bacterial infection. Immune challenges combined with RNA interference targeting NADAR genes led to reduced survival rates and marked necrosis in intestinal tissues, compared to controls exposed to either bacterial infection or dsRNA alone. Together, our results demonstrate that NADAR genes in ladybird beetles were acquired through horizontal gene transfer and contribute to immune defense against bacterial infection.
{"title":"Horizontally transferred NADAR genes contribute to immune defense of ladybird beetles against bacterial infection","authors":"Kun-Yu Yang, Yi-Fei Sun, Yuan-Sen Liang, Hao Li, Meng-Xue Qi, Zhaowei Wang, Luis Carlos Ramos Aguila, Li-Qun Cai, Hao-Sen Li, Hong Pang","doi":"10.1016/j.ibmb.2025.104397","DOIUrl":"10.1016/j.ibmb.2025.104397","url":null,"abstract":"<div><div>Horizontal gene transfer (HGT) is now widely recognized as an important mechanism contributing to host immunity and adaptation. Ladybird beetles, with their diverse diets and habitats, encounter a broad spectrum of microbial threats, making effective immune responses critical for their survival. However, the immune roles of HGT-acquired genes in ladybirds remain largely unexplored. To address this gap, we investigated HGT of a NADAR (NAD- and ADP-ribose-associated) domain-containing gene from microorganisms to insects. Phylogenetic analyses revealed that NADAR genes in ladybird beetles form a well-supported clade nested within a larger group composed primarily of bacterial sequences, providing strong evidence for an HGT origin. Sampling across 69 ladybird species suggests that NADAR genes originated in the Coccinellidae family and were subsequently retained or duplicated across ladybird genomes, indicating their functional importance. Using the ladybird <em>Cryptolaemus montrouzieri</em> as a model, we observed that the expression levels of <em>CmNADAR1</em> and <em>CmNADAR2</em> were significantly upregulated in response to bacterial infection. Immune challenges combined with RNA interference targeting NADAR genes led to reduced survival rates and marked necrosis in intestinal tissues, compared to controls exposed to either bacterial infection or dsRNA alone. Together, our results demonstrate that NADAR genes in ladybird beetles were acquired through horizontal gene transfer and contribute to immune defense against bacterial infection.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104397"},"PeriodicalIF":3.7,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145019854","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 : 2025-09-05DOI: 10.1016/j.ibmb.2025.104398
Merel Braeckman , Lina De Smet , Bart Devreese , Dirk C. de Graaf
This study maps the surfaceome of Apis mellifera hemocytes, the protagonist cells in honey bee cellular immunity. The surfaceome, proteins expressed at the cell surface, is crucial as it determines how cells interact with their microenvironment. Through a combination of proteomic and transcriptomic analyses, 1142 genes encoding cell surface proteins were identified, revealing a high level of diversity. Our analyses identified receptors associated with the major insect immune pathways and proteins previously recognized as hemocyte markers in other invertebrates. Notably, several of the detected genes suggest to encode viral receptors, phagocytosis-related proteins, or proteins involved in hemocyte proliferation. A gene ontology analysis highlighted important functions of the hemocytes. The most prominent cluster was transmembrane receptor protein kinase activity, encompassing over 25 % of the identified terms. Other significant clusters included cell adhesion molecule binding, signalling receptor binding, olfactory receptor activity, and metalloendopeptidase activity. This study suggests several potential honey bee hemocyte markers and establishes a foundation for a novel hemocyte classification based on cell surface markers.
{"title":"Exploration of the hemocyte surfaceome of Apis mellifera by a proteomic and transcriptomic approach","authors":"Merel Braeckman , Lina De Smet , Bart Devreese , Dirk C. de Graaf","doi":"10.1016/j.ibmb.2025.104398","DOIUrl":"10.1016/j.ibmb.2025.104398","url":null,"abstract":"<div><div>This study maps the surfaceome of <em>Apis mellifera</em> hemocytes, the protagonist cells in honey bee cellular immunity. The surfaceome, proteins expressed at the cell surface, is crucial as it determines how cells interact with their microenvironment. Through a combination of proteomic and transcriptomic analyses, 1142 genes encoding cell surface proteins were identified, revealing a high level of diversity. Our analyses identified receptors associated with the major insect immune pathways and proteins previously recognized as hemocyte markers in other invertebrates. Notably, several of the detected genes suggest to encode viral receptors, phagocytosis-related proteins, or proteins involved in hemocyte proliferation. A gene ontology analysis highlighted important functions of the hemocytes. The most prominent cluster was transmembrane receptor protein kinase activity, encompassing over 25 % of the identified terms. Other significant clusters included cell adhesion molecule binding, signalling receptor binding, olfactory receptor activity, and metalloendopeptidase activity. This study suggests several potential honey bee hemocyte markers and establishes a foundation for a novel hemocyte classification based on cell surface markers.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104398"},"PeriodicalIF":3.7,"publicationDate":"2025-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145013575","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 : 2025-08-27DOI: 10.1016/j.ibmb.2025.104396
Zhixin Niu , Huizhen Guo , Dan Li , Yajing Xu , Jianqiu Liu , Yingdan Xiao , Wanshun Li , Amornrat Promboon , Qingyou Xia , Marian R. Goldsmith , Kazuei Mita
Wolbachia is a genus of symbiotic bacteria prevalent in arthropods, with diverse effects on host reproduction and fecundity; however, it is unclear how Wolbachia modulates the host reproductive system. In this study, a novel Wolbachia strain, wSpic, was identified in the Noctuid moth Spodoptera picta and its effect on the reproduction of this host was investigated. We sequenced and annotated the 1,339,720 bp genome of wSpic. We identified a total of five WO phage regions in the genome and found no evidence of any plasmids associated with wSpic. Evolutionary analysis revealed that wSpic belongs to supergroup B and has undergone horizontal transmission between S. picta and Trichogramma pretiosum, a wasp parasitoid of insect eggs. The removal of Wolbachia by antibiotic treatment resulted in significantly decreased fecundity and abnormal development of S. picta ovaries, but no differences in egg hatching rate. An integrated transcriptome and proteome analysis indicated that major molecular pathways for Wolbachia-induced reproduction fitness benefits include its effects on insect juvenile hormone, vitellogenesis, choriogenesis, and nutritional metabolism. Our findings demonstrate that wSpic plays a critical role in promoting ovary development and sustaining fecundity in S. picta hosts.
{"title":"Characterizing the symbiotic relationship between Wolbachia (wSpic) and Spodoptera picta (Lepidoptera: Noctuidae): From genome to phenotype","authors":"Zhixin Niu , Huizhen Guo , Dan Li , Yajing Xu , Jianqiu Liu , Yingdan Xiao , Wanshun Li , Amornrat Promboon , Qingyou Xia , Marian R. Goldsmith , Kazuei Mita","doi":"10.1016/j.ibmb.2025.104396","DOIUrl":"10.1016/j.ibmb.2025.104396","url":null,"abstract":"<div><div><em>Wolbachia</em> is a genus of symbiotic bacteria prevalent in arthropods, with diverse effects on host reproduction and fecundity; however, it is unclear how <em>Wolbachia</em> modulates the host reproductive system. In this study, a novel <em>Wolbachia</em> strain, <em>w</em>Spic, was identified in the Noctuid moth <em>Spodoptera picta</em> and its effect on the reproduction of this host was investigated. We sequenced and annotated the 1,339,720 bp genome of <em>w</em>Spic. We identified a total of five WO phage regions in the genome and found no evidence of any plasmids associated with <em>w</em>Spic. Evolutionary analysis revealed that <em>w</em>Spic belongs to supergroup B and has undergone horizontal transmission between <em>S. picta</em> and <em>Trichogramma pretiosum</em>, a wasp parasitoid of insect eggs. The removal of <em>Wolbachia</em> by antibiotic treatment resulted in significantly decreased fecundity and abnormal development of <em>S. picta</em> ovaries, but no differences in egg hatching rate. An integrated transcriptome and proteome analysis indicated that major molecular pathways for <em>Wolbachia</em>-induced reproduction fitness benefits include its effects on insect juvenile hormone, vitellogenesis, choriogenesis, and nutritional metabolism. Our findings demonstrate that <em>w</em>Spic plays a critical role in promoting ovary development and sustaining fecundity in <em>S. picta</em> hosts.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104396"},"PeriodicalIF":3.7,"publicationDate":"2025-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144917604","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 : 2025-08-23DOI: 10.1016/j.ibmb.2025.104385
Daniel R. Swale , Paul R. Carlier , Maxim Totrov , Jeffrey R. Bloomquist
This study explored how dilution protocols, enzymes, and solvents affected the performance of 2- and 3-phenyl substituted methylcarbamates with varying selectivity for Anopheles gambiae acetylcholinesterase (AgAChE). Protocol A was 100-fold suspension in buffer of a 0.1 M DMSO stock solution, followed by serial buffer dilutions to give declining DMSO in parallel with the inhibitor. Protocol B was identical to A except that the initial stock concentration was 0.01 M. Protocol C entailed DMSO serial dilutions of a 0.1 M DMSO stock solution, followed by suspension of each into buffer giving 0.1 % (v/v) DMSO in all incubates. An. gambiae enzymes from insect homogenates or a recombinant clone generally showed a progressive increase in Hill slope from 0.5 to 1 via protocols A-C, along with increased IC50 values, with the 3-tert-butyl analog (1) epitomizing these effects. In contrast, propoxur displayed no consistent change in inhibition potency of any AChE, regardless of DMSO dilution procedure. DMSO at constant 35 μM or 0.000273 % had a midpoint effect on compound 1 inhibition and displayed competitive inhibition. Time course incubations (10–60 min) over a broad concentration range (10−12 - 10−5 M) of 1 revealed saturable inhibition of high and low potency, with the high potency effect more sensitive to incubation time. In contrast, there was little change of inhibition potency or Hill slope for compound 1 with human AChE, or the AChEs of Drosophila melanogaster and Musca domestica under any DMSO dilution protocol. Moreover, when the dilution protocols were repeated using ethanol as a solvent, little change of inhibition potency or Hill slope was observed with any compound, enzyme, or dilution scheme. These results contradicted the expectation that higher solvent concentration would yield better solubility and more rapid and potent effects of these lipophilic insecticides. Molecular modeling suggests DMSO may be competing with carbamate binding to AgAChE or by stabilizing an allosteric subpocket within AgAChE.
{"title":"Solvent-, enzyme-, and structural-dependence of phenyl-substituted methyl carbamate inhibition of acetylcholinesterase","authors":"Daniel R. Swale , Paul R. Carlier , Maxim Totrov , Jeffrey R. Bloomquist","doi":"10.1016/j.ibmb.2025.104385","DOIUrl":"10.1016/j.ibmb.2025.104385","url":null,"abstract":"<div><div>This study explored how dilution protocols, enzymes, and solvents affected the performance of 2- and 3-phenyl substituted methylcarbamates with varying selectivity for <em>Anopheles gambiae</em> acetylcholinesterase (<em>Ag</em>AChE). Protocol A was 100-fold suspension in buffer of a 0.1 M DMSO stock solution, followed by serial buffer dilutions to give declining DMSO in parallel with the inhibitor. Protocol B was identical to A except that the initial stock concentration was 0.01 M. Protocol C entailed DMSO serial dilutions of a 0.1 M DMSO stock solution, followed by suspension of each into buffer giving 0.1 % (<em>v/v</em>) DMSO in all incubates. <em>An. gambiae</em> enzymes from insect homogenates or a recombinant clone generally showed a progressive increase in Hill slope from 0.5 to 1 via protocols A-C, along with increased IC<sub>50</sub> values, with the 3-<em>tert</em>-butyl analog (<strong>1</strong>) epitomizing these effects. In contrast, propoxur displayed no consistent change in inhibition potency of any AChE, regardless of DMSO dilution procedure. DMSO at constant 35 μM or 0.000273 % had a midpoint effect on compound <strong>1</strong> inhibition and displayed competitive inhibition. Time course incubations (10–60 min) over a broad concentration range (10<sup>−12</sup> - 10<sup>−5</sup> M) of <strong>1</strong> revealed saturable inhibition of high and low potency, with the high potency effect more sensitive to incubation time. In contrast, there was little change of inhibition potency or Hill slope for compound <strong>1</strong> with human AChE, or the AChEs of <em>Drosophila melanogaster</em> and <em>Musca domestica</em> under any DMSO dilution protocol. Moreover, when the dilution protocols were repeated using ethanol as a solvent, little change of inhibition potency or Hill slope was observed with any compound, enzyme, or dilution scheme. These results contradicted the expectation that higher solvent concentration would yield better solubility and more rapid and potent effects of these lipophilic insecticides<em>.</em> Molecular modeling suggests DMSO may be competing with carbamate binding to <em>Ag</em>AChE or by stabilizing an allosteric subpocket within <em>AgA</em>ChE.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104385"},"PeriodicalIF":3.7,"publicationDate":"2025-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144904179","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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