Archives of Insect Biochemistry and Physiology最新文献

{"title":"Comprehensive N-Glycoproteomic Profiling Reveals Unexpected Glycan Diversity and Functional Specialization in the Hemimetabolous Aphid Myzus persicae","authors":"Dongdong Liu,&nbsp;Guy Smagghe","doi":"10.1002/arch.70144","DOIUrl":"10.1002/arch.70144","url":null,"abstract":"<div>\u0000 \u0000 <p>Protein N-glycosylation is a fundamental post-translational modification that regulates protein stability, trafficking, and molecular interactions, yet its diversity and functional significance remain poorly understood in hemimetabolous, phloem-feeding insects. The green peach aphid, <i>Myzus persicae</i>, is a globally important polyphagous pest and vector of numerous plant viruses, making it an ideal system for investigating the physiological and ecological roles of glycosylation. Here, we present the first comprehensive glycoproteomic characterization of <i>M. persicae</i> using high-resolution LC-MS/MS. We identified 43 distinct N-glycan compositions distributed across 446 glycoproteins and 1819 unique glycopeptides. The aphid N-glycome was dominated by oligomannose structures (74%), but also exhibited an unexpectedly high proportion of hybrid (14%) and complex (12%) glycans, contrasting sharply with the &gt; 90% oligomannose dominance reported for holometabolous insects. The most abundant glycans were HexNAc(2)Hex(6) (13%), HexNAc(2)Hex(5) (11%), HexNAc(4)Hex(3) (10%), and the fucosylated paucimannose HexNAc(2)Hex(3)Fuc(1) (10%). Functional enrichment analyses revealed that glycosylation preferentially targets proteins involved in metabolism, detoxification, extracellular processes, and host-plant and virus interactions. Together, these results uncover previously unrecognized N-glycan complexity in a hemimetabolous insect and suggest that expanded hybrid and complex glycosylation may support hemimetabolous development, phloem-feeding specialization, and vector competence. This study establishes a foundational glycoproteomic resource for aphids and provides new insights into glycan-mediated mechanisms with implications for insect physiology, ecology, and the development of glycosylation-targeted pest management strategies.</p></div>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":"121 3","pages":""},"PeriodicalIF":1.9,"publicationDate":"2026-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147503096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ecdysis Triggering Hormone (ETH) Is Vital for Pupation in Henosepilachna vigintioctomaculata","authors":"Lan-Lan Liao,&nbsp;Yi-Kuan Wu,&nbsp;Zhong-Yuan Wang,&nbsp;Juan Peng,&nbsp;Lin Jin,&nbsp;Guo-Qing Li","doi":"10.1002/arch.70146","DOIUrl":"10.1002/arch.70146","url":null,"abstract":"<div>\u0000 \u0000 <p>The initiation of pupal ecdysis behavior by ecdysis-triggering hormone (ETH) has been established in two Lepidopterans and two Dipterans. In Coleoptera <i>Leptinotarsa decemlineata</i>, RNA interference (RNAi) targeting <i>eth</i> impairs pupation. However, whether the RNAi phenotypes result from defects in organ formation or disruption of pupal ecdysis behavior has not been established. Here, we characterized ETH physiological functions during pupation in a potato defoliating ladybird, <i>Henosepilachna vigintioctomaculata</i>. <i>Hveth</i> levels were high just before or right after each molting, it was highly expressed in the trachea-rich tissues, such as dissected epidermis, fat body, and head. RNAi against <i>Hveth</i> impaired pupation of more than 80% of the resultant larvae. They were partially enveloped by the old larval cuticle until death. HE stained slices displayed that new cuticles were well formed under the old ones at the head, pleuron, and pronotum of prothorax. Moreover, adult organs such as compound eyes, mouthparts, and legs were well developed in both the control and the <i>Hveth</i> RNAi groups. Conversely, the freezing sections revealed that no clear separation between the old and the newly formed cuticles in the <i>Hveth</i> RNAi head, in contrast to the control head that showed a visible gap. In addition, the pupal cuticle under the old larval integument was not pigmented. Our findings suggest that ETH initiates pupal ecdysis behavior to shed off old larval exoskeleton and activates pigmentation of pupal cuticle during pupation in <i>H. vigintioctomaculata</i>.</p></div>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":"121 3","pages":""},"PeriodicalIF":1.9,"publicationDate":"2026-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147503064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthetic Peptide Inhibition of Trypsin-Like Proteases in Spodoptera frugiperda (Lepidoptera: Noctuidae): Evaluating the Influence of Gut Microbiota","authors":"Halina Schultz,&nbsp;Daniel Guimarães Silva Paulo,&nbsp;Yaremis Meriño-Cabrera,&nbsp;Rafael Júnior de Andrade,&nbsp;Ian Lucas Batista Santos,&nbsp;Maria Clara Neves Gomes Rodrigues,&nbsp;Geisiane Aparecida Mariano,&nbsp;Milena Godoi Lima,&nbsp;Humberto Josué Ramos de Oliveira,&nbsp;Maria Goreti de Almeida Oliveira","doi":"10.1002/arch.70145","DOIUrl":"10.1002/arch.70145","url":null,"abstract":"<p><i>Spodoptera frugiperda</i> J.E. Smith (Lepidoptera: Noctuidae) is a major agricultural pest whose control has been increasingly challenged by resistance to conventional insecticides. Synthetic protease inhibitors represent promising alternatives; however, the potential modulation of their efficacy by gut microbiota remains poorly understood. Here, we evaluated two rationally designed synthetic tripeptides, GORE1 and GORE2, as inhibitors of trypsin-like digestive proteases and investigated whether gut microbiota disruption alters their inhibitory performance in <i>S. frugiperda</i>. Enzymatic assays revealed competitive inhibition patterns, with <i>K_i</i> values of 1.41 mM for GORE1 and 0.49 mM for GORE2. Most treatments increased apparent <i>K_M</i> values, indicating reduced substrate affinity, whereas GORE2 consistently showed lower <i>K_M</i> values. Despite stronger in vitro affinity (lower <i>K_i</i>), GORE2 did not produce greater biological impairment than GORE1. Microbiota imbalance induced by antibiotic treatment did not significantly alter larval survival or enzymatic inhibition patterns but modulated specific developmental responses, particularly prolonged larval development and reduced body mass under GORE1 exposure. Nutritional indices (ECI and ECD) were significantly reduced in peptide-treated larvae, supporting impaired protein metabolism. These findings demonstrate that synthetic peptides effectively inhibit trypsin-like proteases in <i>S. frugiperda</i> and negatively affect larval nutritional performance. Although microbiota disruption did not modify survival or primary enzymatic inhibition, microbiota-mediated compensatory mechanisms cannot be excluded. Future integrative studies combining microbiome profiling and metabolic analyses will be essential to resolve host–enzyme–microbiota interactions under digestive inhibition.</p>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":"121 3","pages":""},"PeriodicalIF":1.9,"publicationDate":"2026-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12999070/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147479814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Functional Characterization of Spase12 as a Host Factor Contributing to Bombyx mori Nucleopolyhedrovirus Susceptibility in Silkworms","authors":"Nasir Mehmood,&nbsp;Wensheng Yu,&nbsp;Wasim Javaid,&nbsp;Mian Muhammad Awais,&nbsp;Jingchen Sun","doi":"10.1002/arch.70143","DOIUrl":"10.1002/arch.70143","url":null,"abstract":"<div>\u0000 \u0000 <p><i>Bombyx mori</i> is an important economic insect, and its production efficiency is seriously threatened by B. mori nucleopolyhedrovirus (BmNPV). A deeper understanding of host factors exploited during viral infection is crucial for developing disease-resistant silkworm strains. In the current study, we explored the function of <i>Spase12</i>, a subunit of the signal peptidase complex, in the silkworm's antiviral defense. The tissue expression profile showed that <i>Spase12</i> is primarily expressed in the midgut and ovaries, and its expression was significantly upregulated in BmN cells and midgut tissues following BmNPV infection. Additionally, bioinformatics analyses predicted that <i>Spase12</i> is a two-pass endoplasmic reticulum membrane protein, consistent with its role in protein processing and secretion. Moreover, RNAi-mediated knockdown of <i>Spase12</i> significantly suppressed BmNPV replication, as evidenced by decreased <i>VP39</i> expression. On the other hand, overexpression of Spase12 in BmN cells and in vivo experiments using a recombinant baculovirus system confirmed increased viral proliferation and DNA load in the midgut of infected silkworm larvae. Together, these findings established a pro-viral role for <i>Spase12</i> during BmNPV infection, likely hijacked by the virus to facilitate its replication. This study offers novel insights into host–virus interactions in silkworms and highlights <i>Spase12</i> as a potential molecular target for breeding BmNPV-resistant <i>B. mori</i> strains.</p></div>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":"121 3","pages":""},"PeriodicalIF":1.9,"publicationDate":"2026-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147479867","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular Diversity of Gustatory Receptors in Insects: From Structure to Ecological Adaptation","authors":"Rongrong Xu,&nbsp;Jia Zhang,&nbsp;Ting Ge,&nbsp;Ruiqi Song","doi":"10.1002/arch.70142","DOIUrl":"10.1002/arch.70142","url":null,"abstract":"<div>\u0000 \u0000 <p>Gustatory receptors (GRs) are a divergent family of chemoreceptors in insects. By detecting ligands spanning from nutrients to aversive compounds, GRs regulate essential behaviors including feeding, mating, and habitat selection. This multifunctionality makes GRs an indispensable model for studying the sensory-driven evolution paradigm and offers significant translational potential for developing precision targeted attractants or receptor-based pest control strategies. Leveraging insights from phylogenomics, structural biology, and neuroecology, this review systematically examines the molecular diversity and ecological adaptation mechanisms of insect GRs, with the dual objectives of illuminating their evolutionary trajectories and proposing sustainable solutions to address contemporary ecological and agricultural challenges.</p></div>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":"121 3","pages":""},"PeriodicalIF":1.9,"publicationDate":"2026-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147442464","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Snustorr Defines Cuticle Elasticity in Drosophila melanogaster","authors":"Alexander Kovalev,&nbsp;Stanislav Gorb,&nbsp;Bernard Moussian","doi":"10.1002/arch.70140","DOIUrl":"10.1002/arch.70140","url":null,"abstract":"<p>Sir Vincent Wigglesworth proposed in his seminal work on insect cuticle lipids that lipids are involved in cuticle hardening. In brief, in histological experiments, he demonstrated the presence of lipids as a component of cuticulin, a “hard amber material of the insect cuticle.” The molecular evidence for this postulate awaits demonstration. In the present work, we examined the cuticle of larvae lacking the function of the cuticle lipid transporter Snustorr (Snu) by nanoindentation with atomic force microscopy in order to analyze the role of lipids on cuticle hardness in the fruit fly <i>Drosophila melnoagaster</i>. Compared to wild-type larvae, we found that the Young's modulus is 10 times reduced in <i>snu</i> mutant larvae that lack cuticulin at their surface. Thus, this simple result supports the finding of Sir Vincent Wigglesworth that lipids contribute to cuticle hardness in insects.</p>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":"121 3","pages":""},"PeriodicalIF":1.9,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12950260/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147324563","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Histone Deacetylases Are Required for Aedes aegypti Female Reproduction and Embryonic Development","authors":"Sundararajan Balasubramani,&nbsp;Subba Reddy Palli","doi":"10.1002/arch.70139","DOIUrl":"10.1002/arch.70139","url":null,"abstract":"<div>\u0000 \u0000 <p>Histone acetylation and deacetylation are essential for chromatin modifications that regulate various cellular functions. This study identified nine histone deacetylase (HDAC) genes in the yellow fever mosquito, <i>Aedes aegypti</i>, and investigated their roles in female reproduction and embryonic development using RNA interference (RNAi). RNAi-mediated knockdown of HDAC1 (Histone deacetylase 1), HDAC3 (Histone deacetylase 3), HDAC4 (Histone deacetylase 4), and SIRT2 (Sirtuin 2) resulted in severe defects in oocyte development and reduced egg-laying compared with control insects injected with double-stranded RNA (dsRNA) targeting the green fluorescent protein gene (dsGFP). RT-qPCR analysis showed that vitellogenin and its receptor genes are downregulated, leading to suppressed oocyte maturation in mosquitoes injected with dsRNAs targeting HDAC1, HDAC3, HDAC4, and SIRT2 compared with controls. Furthermore, knockdown of HDAC3, HDAC6, and HDAC8 inhibited embryonic development, resulting in reduced hatch rates. These results suggest that HDACs play a vital role in reproduction and embryonic development in <i>Ae. aegypti</i>.</p></div>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":"121 3","pages":""},"PeriodicalIF":1.9,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147324545","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrated In Silico, In Vivo, and Deep Learning Approaches in the Discovery of Novel Candidate Molecules for Aedes aegypti Control","authors":"Herbert Bezerra Leite,&nbsp;Filipe Alves Ribeiro Rodrigues,&nbsp;Luana Beatriz Rocha Silva,&nbsp;Vanessa Costa Santos,&nbsp;Rosalvo F. Oliveira Neto,&nbsp;Edilson B. Alencar Filho","doi":"10.1002/arch.70138","DOIUrl":"10.1002/arch.70138","url":null,"abstract":"&lt;p&gt;The mosquito &lt;i&gt;Aedes aegypti&lt;/i&gt; is a primary vector responsible for transmitting major arboviruses, including dengue, Zika, chikungunya, and yellow fever. Increasing resistance to conventional synthetic insecticides, combined with their well-known environmental drawbacks, underscores the urgent need for more selective, sustainable, and effective strategies for vector control. Chalcones have been previously identified by our research group as a promising chemical class of larvicidal agents, with preliminary evidence for distinct mechanisms of action. More recently, an additional strategy for integrated control of &lt;i&gt;A. aegypti&lt;/i&gt; in its adult stage has emerged through the inhibition of blood feeding, particularly via agonism of neuropeptide Y-like receptor 7 (NPYLR7). In this context, this multi-pronged investigation was conceived as a stage-specific discovery framework addressing distinct biological vulnerabilities of &lt;i&gt;A. aegypti&lt;/i&gt;. Specifically, the study aimed to: evaluate the larvicidal potential of chalcones through integrated in silico and in vivo approaches targeting juvenile hormone transport; apply deep learning–based high-throughput virtual screening (HTVS) as a candidate-prioritization strategy for identifying chemically plausible NPYLR7 agonists associated with blood-feeding inhibition; and finally generate novel NPYLR7-oriented molecular scaffolds using DeSAO (“de novo drugs using Simulated Annealing Optimization)” algorithm as a hypothesis-generating de novo design methodology. These strategies were intentionally pursued as complementary, rather than convergent, discovery axes reflecting the distinct biological requirements of larval and adult mosquito control. Initially, a classical docking-based virtual screening of 1070 chalcones from the PubChem database was conducted on the &lt;i&gt;A. aegypti&lt;/i&gt; juvenile hormone-binding protein (mJHBP), a hemolymph-circulating protein involved in hormonal regulation of larval and adult development. Docking calculations revealed several analogues with favorable predicted binding energies. Three halogenated chalcones were then commercially acquired for experimental larvicidal assays, which identified 4′-chloro-4-methoxychalcone (2c) as the most active compound after 72 h exposure. In parallel, the Machine Learning driven HTVS and the DeSAO workflow independently identified and prioritized novel molecular scaffolds with predicted NPYLR7 agonist activity, generating chemically plausible candidates for subsequent experimental evaluation of blood-meal inhibition in adult mosquitoes. Collectively, the results indicate that halogenated chalcones with moderately sized substituents may serve as promising larvicidal candidates, while HTVS and DeSAO provide complementary, chemically diverse architectures for future evaluation in blood-meal control assays. Taken together, these findings reinforce the value of integrating computational, Machine Learning, and experimental methodologies within a unified p","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":"121 2","pages":""},"PeriodicalIF":1.9,"publicationDate":"2026-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12927534/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147269803","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization and Functional Analysis of Three Cytochrome P450 Monooxygenases in Lasioderma serricorne: Assessing Their Contribution to Lambda-Cyhalothrin Susceptibility Through RNA Interference","authors":"Bin Li,&nbsp;Dao-Shuang Zhu,&nbsp;Yong-Hui Zhang,&nbsp;Chuan-Tao Xu,&nbsp;De-Xin Chen,&nbsp;Mao-Ye Li,&nbsp;Su Liu","doi":"10.1002/arch.70137","DOIUrl":"10.1002/arch.70137","url":null,"abstract":"<div>\u0000 \u0000 <p>In insects, cytochrome P450 monooxygenases (CYPs) play a pivotal role in the detoxification of various insecticides. The cigarette beetle, <i>Lasioderma serricorne</i>, is an economically important pest of stored products, for which pyrethroid insecticides are increasingly used. Nevertheless, the responses and functions of CYPs in <i>L. serricorne</i> under pyrethroid exposure remain poorly understood. In this study, three full-length CYP cDNAs were identified from <i>L. serricorne</i> and designated as <i>CYP6BK29</i>, <i>CYP6SY1</i>, and <i>CYP6TE1</i>. Phylogenetic analysis revealed that these CYPs belong to the CYP3 clan and the CYP6 family. Reverse transcription–quantitative PCR showed that <i>CYP6BK29</i> and <i>CYP6SY1</i> were predominantly expressed in the larval midgut, whereas <i>CYP6TE1</i> transcripts were enriched in the fat body. The expression of <i>CYP6BK29</i> peaked in the third- and fourth-instar larvae, while <i>CYP6SY1</i> was specifically highest in the fourth-instar. In contrast, <i>CYP6TE1</i> showed the highest mRNA levels in adults. Exposure to a sublethal dose (LD₃₀) of lambda-cyhalothrin (LCT) significantly upregulated the mRNA levels of <i>CYP6BK29</i> and <i>CYP6TE1</i>, but not that of <i>CYP6SY1</i>. RNAi-mediated knockdown of <i>CYP6BK29</i> significantly increased the susceptibility of larvae to LCT. In contrast, silencing <i>CYP6SY1</i> or <i>CYP6TE1</i> had no significant effect on larval survival under LCT exposure. These results provide evidence that <i>CYP6BK29</i> contributes to LCT detoxification in <i>L. serricorne</i>, highlighting its potential as a molecular target for novel pest management strategies.</p></div>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":"121 2","pages":""},"PeriodicalIF":1.9,"publicationDate":"2026-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146225318","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DIPA-CRISPR Mediated Knockout of Vermilion Generates a Visible Eye Color Marker for The Band-Legged Ground Cricket Dianemobius nigrofasciatus","authors":"Yuta Shimizu,&nbsp;Kosuke Kataoka","doi":"10.1002/arch.70135","DOIUrl":"10.1002/arch.70135","url":null,"abstract":"<div>\u0000 \u0000 <p>The molecular basis of photoperiodism, by which insects use photoperiodic cues to anticipate seasonal changes and regulate key life-history events such as development, diapause, and reproduction, remains poorly understood. Studies on the molecular mechanisms of photoperiodism in hemimetabolous insects are limited compared with those in holometabolous insects, largely due to the lack of appropriate model organisms. The band-legged ground cricket <i>Dianemobius nigrofasciatus</i> represents a valuable model system because it exhibits clear photoperiodic responses in the maternal induction of embryonic diapause, the wing morph, and the rate of nymphal development. With the recent availability of the <i>D. nigrofasciatus</i> genome sequence, the establishment of effective genome-editing methods and reliable marker genes is expected to promote functional genomic analyses. In this study, we aimed to establish a direct parental (DIPA)-CRISPR genome-editing approach and evaluate the utility of <i>vermilion</i> (<i>Dn-v</i>), a gene involved in ommochrome synthesis, as a visible eye color marker for mutant screening. Cas9 ribonucleoprotein complexes were injected into females 3–5 days after adult emergence, during the vitellogenic stage, successfully yielding <i>Dn-v</i> knockout mutants. These mutants had white compound eyes throughout development, with pigmentation reaching a vermilion color about 20 days after adult emergence. We further examined the photoperiodic response associated with maternal diapause induction in knockout mutants. Similar to the wild-type, knockout mutants exhibited low and high diapause incidence under long-day and short-day conditions, respectively. Our results demonstrate that DIPA-CRISPR is an effective genome-editing method in <i>D. nigrofasciatus</i> and that <i>Dn-v</i> serves as a practical and reliable marker gene. The establishment of these genomic tools provides a foundation for future functional analyses aimed at elucidating the molecular basis of photoperiodism in hemimetabolous insects.</p></div>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":"121 2","pages":""},"PeriodicalIF":1.9,"publicationDate":"2026-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146225361","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}