Yuchen Zhao , Pei Wang , Yalan Cheng , Siyu Tian , Junfeng Zuo , Qianguang Fang , Chenxue Ling , Subba Reddy Palli , Xien Chen
{"title":"A testis-specific heat shock protein gene plays a critical role in male fertility in the fall armyworm","authors":"Yuchen Zhao , Pei Wang , Yalan Cheng , Siyu Tian , Junfeng Zuo , Qianguang Fang , Chenxue Ling , Subba Reddy Palli , Xien Chen","doi":"10.1016/j.ibmb.2025.104332","DOIUrl":"10.1016/j.ibmb.2025.104332","url":null,"abstract":"<div><div>Heat shock proteins (Hsps) are critical for stress responses and multiple physiological processes, yet their function in insect reproduction remains underexplored. In this study, we identified 36 Hsp genes in the fall armyworm (<em>Spodoptera frugiperda</em>), of which LOC118269601 (designated as <em>SfHsp68A</em>) exhibited robust testis-specific expression, peaking during pupal testis development. CRISPR/Cas9-mediated knockout of <em>SfHsp68A</em> resulted in recessive male sterility, significantly reduced testis size, sperm counts, and egg hatch rates. Transcriptomic analysis of mutant testes revealed substantial downregulation of genes linked to mitochondrial function, oxidative phosphorylation, energy metabolism, and reactive oxygen species (ROS) metabolism. These results demonstrate that SfHsp68A is essential for male fertility in <em>S. frugiperda,</em> likely by maintaining mitochondrial integrity and metabolic homeostasis during spermatogenesis. These studies reveal that testis-specific Hsps function in male fertility and highlight <em>SfHsp68A</em> as a potential genetic target for developing sterile insect technology, offering an environmentally sustainable strategy for controlling this globally invasive pest.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"181 ","pages":"Article 104332"},"PeriodicalIF":3.2,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144166977","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}
Mariana Lizbeth Jiménez-Martínez , Gerardo de Jesús Trujillo-Rodriguez , María de Lourdes Ramírez-Ahuja , Olga Karina Villanueva-Segura , Patricio Adrián Zapata-Morin , Griffith Samuel Lizarraga , Laura E. Martinez-de-Villarreal , Margarita L. Martinez-Fierro , Ivan Delgado-Enciso , Jose Ignacio Gonzalez-Rojas , Adriana E. Flores , Antonio Guzman-Velasco , Iram Pablo Rodriguez-Sanchez
{"title":"Amino acid and acylcarnitine profile of Aedes aegypti (L) post-blood feeding, after permethrin exposure, and during the life cycle","authors":"Mariana Lizbeth Jiménez-Martínez , Gerardo de Jesús Trujillo-Rodriguez , María de Lourdes Ramírez-Ahuja , Olga Karina Villanueva-Segura , Patricio Adrián Zapata-Morin , Griffith Samuel Lizarraga , Laura E. Martinez-de-Villarreal , Margarita L. Martinez-Fierro , Ivan Delgado-Enciso , Jose Ignacio Gonzalez-Rojas , Adriana E. Flores , Antonio Guzman-Velasco , Iram Pablo Rodriguez-Sanchez","doi":"10.1016/j.ibmb.2025.104328","DOIUrl":"10.1016/j.ibmb.2025.104328","url":null,"abstract":"<div><div><em>Aedes aegypti</em>, a major vector for arboviruses, requires effective control strategies due to rising insecticide resistance. This study quantified 12 amino acids and 24 acylcarnitines in two strains of <em>Ae. aegypti</em> (New Orleans, susceptible; Apodaca, resistant) using tandem mass spectrometry (MS/MS) across three approaches: 1) life cycle stages, 2) fasting and post-blood feeding (24, 48, 72 h), and 3) before and after permethrin exposure. In the life cycle, alanine (Ala) was highest in adult males (40.3 %), arginine (Arg) peaked in adult females (18.35 %), and tyrosine (Tyr) reached 14.74 % in larvae. The acylcarnitine C0 (free carnitine) peaked in larvae stage 4 (94.21 %), while C2 (acetyl carnitine) was highest in adult females (47.68 %). Post-blood feeding, Ala peaked at 72 h (43.34 %), proline (Pro) at 48 h (20.63 %), and Arg decreased from 0 h (18.35 %) to 72 h (11.15 %). Acylcarnitines C0 and C2 showed a decline post-feeding, indicating fatty acid mobilization. After permethrin exposure, resistant mosquitoes exhibited higher Ala (51.7 %) and C2 (57.9 %) levels, suggesting increased metabolic activity for detoxification. In contrast, susceptible mosquitoes displayed less variability in amino acid and acylcarnitine levels. These results provide valuable insights into the metabolic adaptations of <em>Ae. aegypti</em>, indicating the potential for targeting amino acid and acylcarnitine pathways to improve vector control strategies, particularly in the context of rising insecticide resistance.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"181 ","pages":"Article 104328"},"PeriodicalIF":3.2,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144140985","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Identification of biliverdin as a pigment involved in the seasonal variation of green lacewing body colour and the enzymes involved in its metabolism","authors":"Shion Abe, Yuka Matsubara, Kanako Ogawa, Yuki Nakamura, Satoshi Yamauchi, Hisashi Nishiwaki","doi":"10.1016/j.ibmb.2025.104330","DOIUrl":"10.1016/j.ibmb.2025.104330","url":null,"abstract":"<div><div>The green lacewing, <em>Chrysoperla nipponensis</em>, is an insect that undergoes complete metamorphosis, and its body colour changes depending on the developmental stage; adults and larvae are greenish and brownish, respectively. In addition, the body colour of some adults changes to brown in winter, suggesting that environmental factors influence pigment biosynthesis in this insect. However, the underlying colour regulation systems remain unknown. In this study, we purified the pigments that produce the body colour of green adult lacewings and identified biliverdin via MS/MS analyses. We therefore elucidated the mRNA and amino acid sequences of biliverdin-related biosynthetic enzymes, such as haem oxygenase (HO) and biliverdin reductase (BR), in <em>C. nipponensis</em>. By expressing recombinant BR in <em>E. coli</em>, we investigated the ability of BR to metabolize biliverdin. In addition, via RNAi techniques, we examined whether the suppression of these enzymes changed body colour. Finally, we quantified the expression levels of the mRNAs encoding these enzymes and compared those in green adults reared under normal conditions with those in brown adults reared under winter-mimicking conditions. The results suggest that not only biliverdin but also other green pigments contribute to the green body colour of adult lacewings.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"181 ","pages":"Article 104330"},"PeriodicalIF":3.2,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144126333","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}
Hui-Jie Wu , Jia-Peng Yang , Wen-Jing Ma , Zheng-Hao Li , Hong-Yu Feng , Zhang-Nv Yang , Hai-Jun Xu
{"title":"A CRISPR/Cas9-induced point mutation on the GABA receptor subunit RDL confers high resistance to phenylpyrazole insecticides in the rice planthopper Laodelphax striatellus","authors":"Hui-Jie Wu , Jia-Peng Yang , Wen-Jing Ma , Zheng-Hao Li , Hong-Yu Feng , Zhang-Nv Yang , Hai-Jun Xu","doi":"10.1016/j.ibmb.2025.104327","DOIUrl":"10.1016/j.ibmb.2025.104327","url":null,"abstract":"<div><div>The Delphacidae planthopper <em>Laodelphax striatellus</em> (Order: Hemiptera) is one of the most damaging insect pests of rice crops in Asia. The phenylpyrazole insecticide fipronil was introduced in China in the mid-1990s to control these pests, but its widespread use has led to the development of high levels of resistance. Field sampling coupled with <em>in vitro</em> assays indicated that an A2′N-point mutation in the gamma-aminobutyric acid receptor RDL has been linked to fipronil resistance; however, genetic evidence supporting this association has been lacking. Here, we generated an A2′N-point mutation (<em>LsRDL</em><sup><em>A/N</em></sup>) in <em>L. striatellus</em> using CRISPR/Cas9-mediated homologous recombination. Bioassay revealed that the <em>LsRDL</em><sup><em>A/N</em></sup> mutants conferred a 1211-fold higher resistance to fipronil compared to wild-type planthoppers. Furthermore, these mutants also showed significant resistance (>1000-fold) to ethiprole but not to imidacloprid, demonstrating that the A2′N mutation confers resistance specific to phenylpyrazole insecticides. However, the <em>LsRDL</em><sup><em>A/N</em></sup> mutants displayed reduced fecundity, lower egg hatching rates, and prolonged egg and nymph stages, indicating that fipronil resistance carries a substantial fitness cost. These findings enhance our understanding of the genetic mechanisms underlying phenylpyrazole resistance and provide valuable insights for developing effective resistance management strategies for planthoppers.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"181 ","pages":"Article 104327"},"PeriodicalIF":3.2,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144124776","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}
Yunlong Zou, Aijun Ye, Meixin Dong, Yuhao Zhou, Wentao Wu, Yuxia Tang, Hai Hu, Fangyin Dai, Xiaoling Tong
{"title":"Silkworm mutagenesis using a ribonucleoprotein-based CRISPR/Cas12a system.","authors":"Yunlong Zou, Aijun Ye, Meixin Dong, Yuhao Zhou, Wentao Wu, Yuxia Tang, Hai Hu, Fangyin Dai, Xiaoling Tong","doi":"10.1016/j.ibmb.2025.104329","DOIUrl":"https://doi.org/10.1016/j.ibmb.2025.104329","url":null,"abstract":"<p><p>The development of highly efficient genome editing tools has revolutionized developmental biology and genetic studies in silkworm. Although methods based on CRISPR/Cas9 are currently popular, the Cas12a system has emerged as a promising option. However, it has not yet been applied to target the silkworm genome in vivo, and its activity in silkworm has not yet been characterized. In this study, we established a ribonucleoprotein-based CRISPR/Cas12a system, and compared it to the CRISPR/Cas9 system using 19 crRNA and 17 sgRNAs to target three different genes in vivo. Although Cas12a generates mutants less efficiently than Cas9, we used it successfully to generate transmissible indels, and demonstrated its application by targeting the FibH and mp genes to produce mutants with the expected phenotypes. We also assessed the influence of temperature (37 °C vs. 25 °C) on Cas12a activity, and demonstrated that the effects are target dependent. In summary, we have established a ribonucleoprotein-based CRISPR/Cas12a system in silkworm that offers a practical alternative to CRISPR/Cas9 and extends the genome editing tool box available for silkworm research.</p>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":" ","pages":"104329"},"PeriodicalIF":3.2,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144118503","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Mini-review: Aspects of cuticle formation and structure advanced by studies in Nilaparvata lugens.","authors":"Chuan-Xi Zhang, Bernard Moussian","doi":"10.1016/j.ibmb.2025.104326","DOIUrl":"https://doi.org/10.1016/j.ibmb.2025.104326","url":null,"abstract":"<p><p>A hallmark of insects is their cuticle that protects them against predators and parasites, prevents desiccation and xenobiotic penetration and, finally, serves locomotion as an exoskeleton. In the last two decades, tremendous efforts have yielded a plethora of molecular and histological data that have boosted our understanding of the mechanisms of cuticle formation and function. As the factors involved in cuticle formation and function are to a large extent specific to insects, these data may be used in intelligent pest management strategies. The brown planthopper Nilaparvata lugens is a major rice pest in Asia. Cuticle research in this species has been following findings in other insect species recapitulating most collected results. There are, in addition, several important data in N. lugens that advanced our understanding in cuticle biology in insects generally. For instance, it was demonstrated in the brown planthopper that a chitin-binding protein, namely NlugCpr47 is needed for organ shaping. Moreover, based on genetic data, it was hypothesised that cuticular lipids may be organised in layers depending on the size of the molecules. Finally, pore canals, the nanotubes system that is used as a transport route for cuticle lipid deposition, were visualised in 3D with high resolution for the first time in an insect. These examples, reviewed in this article, underline the importance of a detailed molecular and histological approach in non-model insects for a better understanding of cuticle biology in general.</p>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":" ","pages":"104326"},"PeriodicalIF":3.2,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144085550","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}
Kai Chen , Ling Ye , Ye Yu , Peilin Guo , Anjiang Tan
{"title":"Sex-biased fertility degeneration induced by depletion of an auxiliary piRNA-pathway factor Qin in Bombyx mori","authors":"Kai Chen , Ling Ye , Ye Yu , Peilin Guo , Anjiang Tan","doi":"10.1016/j.ibmb.2025.104319","DOIUrl":"10.1016/j.ibmb.2025.104319","url":null,"abstract":"<div><div>The PIWI-interacting RNA (piRNA) pathway is the major defense system for transposable elements (TEs) silencing in animal gonads, maintaining genomic integrity of germ cells and ensuring proper gametogenesis. An the piRNA-pathway factor, <em>Qin</em>, has been reported to participate in piRNA biogenesis in the lepidopteran model insect, <em>Bombyx mori</em>. Nevertheless, the physiological functions of <em>Qin</em> remain to be characterized. Here we demonstrated that Qin plays important roles in silkworm gonad development of both sexes. <em>BmQin</em> was predominantly expressed in gonads. Immunofluorescent staining revealed that BmQin is localized in the cytoplasm of both germ cells and somatic cells in gonads. Depletion of <em>BmQin</em> via CRISPR/Cas9 system induceed complete sterile in males, and partial sterile in females. Notably, mutants displayed severe defects in gonad development and gametogenesis. RNA-seq analysis revealed that the piRNA pathway was dysregulated in mutant gonads. In addition, apoptosis was significantly enhanced in mutant gonads. Our study revealed the physiological functions of <em>BmQin</em> in silkworm fertility and its auxiliary roles in the piRNA pathway in both male and female gonads.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"181 ","pages":"Article 104319"},"PeriodicalIF":3.2,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143928346","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}
Ting-Ting Yang , Yu Ma , Zheng Dai , Ying Li , Ji-Xiang Wang , Teng-Fei Bai , Merid Negash Getahun , George F.O. Obiero , Shuang-Lin Dong , Jin Zhang , Qi Yan
{"title":"Molecular mechanisms based on peripheral level of vanillin recognition in Orthaga achatina (Lepidoptera: Pyralidae)","authors":"Ting-Ting Yang , Yu Ma , Zheng Dai , Ying Li , Ji-Xiang Wang , Teng-Fei Bai , Merid Negash Getahun , George F.O. Obiero , Shuang-Lin Dong , Jin Zhang , Qi Yan","doi":"10.1016/j.ibmb.2025.104317","DOIUrl":"10.1016/j.ibmb.2025.104317","url":null,"abstract":"<div><div><em>Orthaga achatina</em> (Lepidoptera Pyralidae) is a specialist pest of the camphor tree <em>Cinnamomum camphora</em>. Vanillin is a volatile compound found in many plants, and its effects on insects can be either attractive or repellent, depending on the species. However, the behavioral response of <em>Orthaga achatina</em> to vanillin, a volatile compound emitted by camphor trees, remains unexplored. In this study, we found that vanillin attracts both male and female <em>O. achatina</em> adults. Fluorescence competitive binding assays further revealed that among the five odorant-binding proteins (OBPs) highly expressed in both male and female antennae, OachOBP7 exhibited the most prominent binding affinity with vanillin. Furthermore, by employing the Xenopus oocyte expression and two-electrode voltage clamp recording system (XOE-TEVC) to conduct a functional characterization of 40 ORs, vanillin was the optimal ligand for OachOR7 among all tested ligands. In addition, with the 3D structure modeling and molecular docking techniques, it was revealed that OachOR7 displayed a relatively high binding affinity (−5.5 kcal/mol), and Gln84 and Asn189 were predicted to be key amino acid residues for binding vanillin. Finally, the two amino acids were verified by site-specific mutagenesis followed by XOE-TEVC, showing that the binding ability of OR7 to vanillin was significantly reduced to 9.23 × 10<sup>−2</sup> μM after the mutation of two amino acids. This study demonstrate vanillin's behavioral attraction to <em>O. achatina</em> and reveal its molecular basis, offering new possibilities for targeted pest management using this compound.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"180 ","pages":"Article 104317"},"PeriodicalIF":3.2,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143891843","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}
Wei Dong , Xinyi Wang , Zining Cheng , Bernard Moussian , Jianzhan Zhang , Xu-Bo Zhang
{"title":"HiHR4 regulates chitin metabolism during molting of Hermetia illucens","authors":"Wei Dong , Xinyi Wang , Zining Cheng , Bernard Moussian , Jianzhan Zhang , Xu-Bo Zhang","doi":"10.1016/j.ibmb.2025.104318","DOIUrl":"10.1016/j.ibmb.2025.104318","url":null,"abstract":"<div><div>The black soldier fly (<em>Hermetia illucens</em>) as a model bio-converter insect, is increasingly used in feed formulation and as a fertilizer source. Its growth rate and biomass yield are influenced by molting, a process involving the remodeling of chitinous structures. In this study, we identified a <em>HiHR4</em> gene in <em>Hermetia illucens</em> and report its function in larval growth and molting. RT-qPCR results show that <em>HiHR4</em> expression is stable during the third instar larval stage but peaks sharply prior to molting. Knockdown of <em>HiHR4</em> by RNA interference (RNAi) in larvae results in molting delay. The ds<em>HiHR4-</em>injected larvae display slow growth. Consistently, integument sectioning and chitin staining showed that <em>HiHR4</em> knockdown inhibits both the degradation of the old cuticle and the formation of new cuticle. In addition, <em>HiHR4</em> and chitin metabolism-related genes are induced by the steroid hormone 20-hydroxyecdysone (20E), indicating their responsiveness to 20E signaling. Moreover, <em>HiHR4</em> knockdown significantly suppresses the expression of chitin synthesis genes (<em>HiHex, HiG6PI, HiCHS1</em>) and chitin degradation genes (<em>HiCht10</em>). RNAi against <em>HiCHS1</em> or <em>HiCht10</em> disrupts larval molting and attenuates growth. Taken together, these findings suggest that <em>HiHR4</em> is a key regulator of chitin metabolism, ensuring proper molting and promoting biomass production in <em>H. illucens</em>.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"181 ","pages":"Article 104318"},"PeriodicalIF":3.2,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143906452","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Role of histone methylation in insect development: KMT5A regulates ecdysteroid biosynthesis during metamorphosis of Tribolium castaneum","authors":"Yaoyu Jiao , Karthi Sengodan, Jiasheng Chen , Subba Reddy Palli","doi":"10.1016/j.ibmb.2025.104316","DOIUrl":"10.1016/j.ibmb.2025.104316","url":null,"abstract":"<div><div>Methylation levels of core histones play important roles in the regulation of gene expression and impact animal development. However, the methyltransferases and demethylases that determine histone methylation levels remain largely unexplored in insects. Most of our current understanding of histone methylation comes from mammalian studies. In this study, we first identified potential histone methyltransferases and demethylases encoded in the genome of the red flour beetle <em>Tribolium castaneum</em>. The function of these histone methylation enzymes in the metamorphosis was investigated by knocking down genes coding for these enzymes using RNA interference (RNAi). Our results showed that a lysine methyltransferase, KMT5A, plays a critical role in <em>T. castaneum</em> metamorphosis by regulating the biosynthesis of ecdysteroids. Treating <em>KMT5A</em>-knockdown larvae with 20 hydroxyecdysone can partially rescue <em>T. castaneum</em> pupation. Western blot analysis showed that KMT5A catalyzes H4K20 mono-methylation. However, further studies suggest that KMT5A may regulate <em>T. castaneum</em> pupation through mechanisms independent of H4K20 methylation. These data uncovered the roles of histone methylation enzymes in <em>T. castaneum</em> metamorphosis and KMT5A as a critical regulator of ecdysteroid biosynthesis.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"180 ","pages":"Article 104316"},"PeriodicalIF":3.2,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143881836","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}