Insect Biochemistry and Molecular Biology最新文献_第6页

Replication and transcription of eccDNAfib−L, an extrachromosomal circular DNA in the silk gland of silkworm, Bombyx mori 家蚕蚕丝腺染色体外环状DNA eccDNAfib-L的复制和转录。

IF 3.7 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2025-10-17 DOI: 10.1016/j.ibmb.2025.104424

Xinyu Tong , Chao Lei , Mei Yin , Qunnan Qiu , Xiaolong Hu , Min Zhu , Chengliang Gong

Extrachromosomal circular DNA (eccDNA) plays critical roles in gene regulation and genome plasticity. Some eccDNAs, known as microDNAs, are less than 1000 bp in length and are capable of autonomous transcription. However, whether microDNAs can replicate remains unclear. We previously identified an eccDNA, eccDNA^fib−L, which is 542 bp in size (Chromosome 14: 9,692,083–9,692,624nt) in the silk gland of Bombyx mori. While eccDNA^fib−L promotes the expression of the fib-L gene, its replication and transcription capabilities remain unknown. In this study, we revealed that eccDNA^fib−L exists in two forms: double-stranded (eccDNA^fib−L-ds) and single-stranded forms (eccDNA^fib−L-ss(+), eccDNA^fib−L-ss(−)). Both double-stranded and single-stranded eccDNA^fib−L exhibit replication capability, with its replication mechanism potentially analogous to that of mitochondrial DNA. Furthermore, eccDNA^fib−L-ds undergoes rolling circle transcription without termination. Transcription of eccDNA^fib−L-ds utilizes the antisense (−) strand as the template, with the transcription direction consistent with that of the fib-L gene. Structural analysis indicated that the GC nucleotides at position 505–506 nt (with the first nucleotide at the junction site of eccDNA^fib−L-ds defined as position 1) serve as the transcription start site, which was validated by the absence of transcripts following the mutation of GC to TG/TA. The transcription process of eccDNA^fib−L-ds is dependent on RNA polymerase Ⅱ and multiple regulatory factors. These findings demonstrate that eccDNA^fib−L can replicate and transcribe, enhancing our understanding of its role in regulating silk protein synthesis and expanding our knowledge of the functions of eccDNAs.

染色体外环状DNA在基因调控和基因组可塑性中起着至关重要的作用。一些eccdna，被称为微dna，长度小于1000 bp，能够自主转录。然而，微dna是否能够复制尚不清楚。我们先前在家蚕蚕丝腺中发现了一个eccDNAfib-L，全长542 bp（染色体14:9,692,083-9,692,623 nt）。虽然eccDNAfib-L促进了fib-L基因的表达，但其复制和转录能力尚不清楚。在这项研究中，我们发现eccDNAfib-L以两种形式存在：双链形式（eccDNAfib-L-ds）和单链形式(eccDNAfib-L-ss（+), eccDNAfib-L-ss(-)）。双链和单链eccDNAfib-L都具有复制能力，其复制机制可能类似于线粒体DNA。此外，eccDNAfib-L-ds进行滚动循环转录而不终止。eccDNAfib-L-ds的转录以反义（-）链为模板，转录方向与fib-L基因一致。结构分析表明，位于505-506 nt位置的GC核苷酸（eccDNAfib-L-ds连接位点的第一个核苷酸定义为位置1）作为转录起始位点，通过GC突变为TG/TA后缺乏转录物证实了这一点。eccDNAfib-L-ds的转录过程依赖于RNA聚合酶Ⅱ和多种调控因子。这些发现表明eccDNAfib-L可以复制和转录，增强了我们对其在调节丝蛋白合成中的作用的理解，并扩展了我们对eccdna功能的认识。

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引用次数: 0

Drosophila proteasome subunit Rpn8 controls IMD pathway activation via PGRP-SC2 degradation 果蝇蛋白酶体亚基Rpn8通过PGRP-SC2降解控制IMD途径的激活。

IF 3.7 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2025-10-11 DOI: 10.1016/j.ibmb.2025.104422

Mingfei Wu , Umar Ali , Sen Ding , Chuchu Zhang , Yiheng Jin , Xinyan Cao , Muhammad Abdul Rehman Saeed , Yangyang Zhu , Muhammad Usama , Badar Raza , Qingshuang Cai , Shanming Ji

The ubiquitin-proteasome system (UPS) is a highly conserved protein degradation pathway in insects, playing crucial roles in cellular homeostasis, signal transduction, and various biological processes. Here, we demonstrate that the Drosophila melanogaster (fruit fly) regulatory particle non-ATPase 8 (Rpn8), a key subunit of the 26 S proteasomal cap, is indispensable for fly survival upon bacterial infections. Through targeted genetic knockdown, we observed that Rpn8-silenced flies exhibited decreased expressions of several antimicrobial peptides upon bacterial challenges, concomitant with impaired bacterial clearances. Further proteomic and biochemical analyses revealed that Rpn8 controls the proteasome-mediated degradation of peptidoglycan recognition protein SC2 (PGRP-SC2), a typical amidase that antagonizes immune deficiency (IMD) signaling. Silencing of PGRP-SC2 prevented the functional role of Rpn8 in controlling the fly antibacterial immune defense, suggesting that Rpn8 mediates Drosophila innate immunity in a PGRP-SC2-dependent manner. Intriguingly, overexpression of Bombyx mori (silkworm) Rpn8 rescued the immune defects in Rpn8 RNAi flies after bacterial infections. Collectively, our findings not only establish Rpn8 as a novel immune modulator that bridges UPS-mediated protein degradation with innate immune defense in Drosophila but also offer insights into conserved mechanisms of host-pathogen interactions in other insect species.

泛素-蛋白酶体系统（ubiquitin-proteasome system， UPS）是昆虫体内高度保守的蛋白质降解途径，在细胞稳态、信号转导和多种生物过程中起着至关重要的作用。在这里，我们证明果蝇的调节颗粒非atp酶8 （Rpn8）是26S蛋白酶体帽的一个关键亚基，对于细菌感染后果蝇的生存是必不可少的。通过靶向基因敲低，我们观察到rpn8沉默的果蝇在细菌攻击时表现出几种抗菌肽的表达减少，同时细菌清除能力受损。进一步的蛋白质组学和生化分析表明，Rpn8控制蛋白酶体介导的肽聚糖识别蛋白SC2 （PGRP-SC2）的降解，PGRP-SC2是一种典型的抗免疫缺陷（IMD）信号的酰胺酶。PGRP-SC2的沉默阻止了Rpn8在控制果蝇抗菌免疫防御中的功能作用，提示Rpn8以PGRP-SC2依赖的方式介导果蝇先天免疫。有趣的是，家蚕Rpn8的过表达挽救了细菌感染后Rpn8 RNAi蝇的免疫缺陷。总的来说，我们的研究结果不仅确立了Rpn8作为一种新的免疫调节剂，在果蝇中架起了ups介导的蛋白质降解与先天免疫防御之间的桥梁，而且还为其他昆虫物种中宿主-病原体相互作用的保守机制提供了见解。

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引用次数: 0

A conserved Piezo–mRPL2/Jun axis controls reproduction via mitochondrial bioenergetics 保守的Piezo-mRPL2/Jun轴通过线粒体生物能量学控制生殖。

IF 3.7 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2025-10-10 DOI: 10.1016/j.ibmb.2025.104417

Jiangwen Luo , Yaru Lu , Zhiruo Zhang , Qian Zhang, Yunzhu Shang, Yajie Yuan, Xiaohui Duan, Long Zhang, Yuxia Tang, Hai Hu, Xiaoling Tong, Fangyin Dai

Piezo channels are essential mechanosensors, but their roles in reproduction remain poorly understood. Here, we identify a critical function for BmPiezo in silkworm fertility and embryonic diapause, mediated through a conserved downstream regulatory axis. CRISPR/Cas9-mediated knockout of BmPiezo resulted in male sterility and loss of embryonic diapause, accompanied by dysregulation of ribosome-associated genes and oxidative phosphorylation, and reduced ATP production. We identified mitochondrial ribosomal protein L2 (BmmRPL2) and transcription factor BmJun as key downstream effectors, required for spermatogenesis and diapause, respectively. Homozygous disruption of either gene caused embryonic lethality. Cross-species expression analysis revealed strong expression of Piezo2 and mRPL2 in early human spermatids. Remarkably, transgenic expression of murine Piezo2 in BmPiezo−/−silkworms reactivated the Piezo–mRPL2/Jun axis, restored ATP levels, and rescued reproductive defects. These findings define a conserved pathway that connects mechanical sensing to mitochondrial bioenergetics in reproductive regulation, advancing our understanding of fertility mechanisms across species.

压电通道是必不可少的机械传感器，但它们在繁殖中的作用仍然知之甚少。在这里，我们确定了BmPiezo在家蚕生育和胚胎滞育中的关键功能，通过保守的下游调节轴介导。CRISPR/ cas9介导的敲除BmPiezo导致雄性不育和胚胎滞育丧失，并伴有核糖体相关基因和氧化磷酸化的失调，以及ATP的产生减少。我们发现线粒体核糖体蛋白L2 （BmmRPL2）和转录因子BmJun分别是精子发生和滞育所需的关键下游效应物。任何一个基因的纯合破坏都会导致胚胎死亡。跨种表达分析显示，Piezo2和mRPL2在早期人类精子中有较强的表达。值得注意的是，转基因小鼠Piezo2在BmPiezo-/-家蚕体内的表达可重新激活piezomrpl2 /Jun轴，恢复ATP水平，并挽救生殖缺陷。这些发现定义了一条保守的途径，将机械传感与生殖调节中的线粒体生物能量学联系起来，促进了我们对物种生育机制的理解。

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引用次数: 0

Bombyx mori nucleopolyhedrosis virus-derived circular RNAs with protein-coding potential facilitate viral replication 家蚕核多角体病毒衍生的环状rna具有蛋白质编码潜能，可促进病毒复制。

IF 3.7 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2025-10-09 DOI: 10.1016/j.ibmb.2025.104420

Xing Zhang , Zeen Shen , Yani Kang , Wenbin Yu , Xiaoyan Du , Qian Teng , Zihan He , Chengliang Gong , Xiaolong Hu

Circular RNAs (circRNAs) are renowned for their exceptional stability and have been increasingly recognized for their dual roles in pro- and antiviral responses during host-virus interactions. In this study, we investigated the functional landscape of circular RNAs (circRNAs) during infection with Bombyx mori nucleopolyhedrosis virus (BmNPV), a model baculovirus system. Our comprehensive analysis revealed that hundreds of host-derived circRNAs are dynamically regulated upon BmNPV infection, while the virus itself generates numerous viral circRNAs (vcircRNAs) via back-splicing. Using a combination of advanced experimental approaches, we validated the existence of multiple cellular and viral circRNAs. Among these, we characterized vcircRNA-390, a BmNPV-encoded circRNA harboring a small open reading frame (ORF) and four viral internal ribosome entry sites (IRESs). Remarkably, vcircRNA-390 serves as a template for the translation of an 81-amino acid viral peptide (VSP81). Functional studies demonstrated that both vcircRNA-390 and VSP81 significantly enhance viral replication. Mechanistically, we provide evidence that VSP81 likely targets the host RNA interference (RNAi) pathway, a major antiviral defence system, to promote viral immune evasion. While these findings establish vcircRNA-390 as a novel proviral factor in insect-virus interactions, the detailed molecular mechanisms by which VSP81 modulates the RNAi machinery remain to be fully elucidated. Our work not only expands the understanding of viral circRNA biology but also reveals a new dimension of the host-pathogen conflict, wherein BmNPV exploits circRNA-mediated translation to subvert antiviral defences.

环状rna （circRNAs）以其卓越的稳定性而闻名，并因其在宿主-病毒相互作用期间的亲抗病毒反应中的双重作用而日益得到认可。在这项研究中，我们研究了环状rna （circRNAs）在感染家蚕核多角体病毒（BmNPV）时的功能景观。我们的综合分析显示，数百种宿主衍生的环状rna在BmNPV感染时受到动态调节，而病毒本身通过反剪接产生许多病毒环状rna （vcircRNAs）。结合先进的实验方法，我们验证了多种细胞和病毒环状rna的存在。其中，我们鉴定了vcircRNA-390，这是一种bmnpv编码的环状rna，含有一个小的开放阅读框（ORF）和四个病毒内部核糖体进入位点（IRESs）。值得注意的是，vcircRNA-390可作为81个氨基酸的病毒肽（VSP81）翻译的模板。功能研究表明，vcircRNA-390和VSP81均能显著增强病毒复制。在机制上，我们提供的证据表明，VSP81可能靶向宿主RNA干扰（RNAi）途径，一个主要的抗病毒防御系统，以促进病毒免疫逃避。虽然这些发现确定了vcircRNA-390是昆虫-病毒相互作用中的一个新的原病毒因子，但VSP81调节RNAi机制的详细分子机制仍有待充分阐明。我们的工作不仅扩展了对病毒circRNA生物学的理解，而且揭示了宿主-病原体冲突的一个新维度，其中BmNPV利用circRNA介导的翻译来破坏抗病毒防御。

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引用次数: 0

Assessment of RNA interference effectiveness mediated by siRNA sequences and structures in Drosophila S2 cells 果蝇S2细胞中siRNA序列和结构介导的RNA干扰有效性评估。

IF 3.7 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2025-10-09 DOI: 10.1016/j.ibmb.2025.104418

Hyun-Soo Kim , Huipeng Pan , Subba Reddy Palli , June-Sun Yoon

Extensive research has clarified the mechanisms of RNA interference (RNAi) in insects. Although double-stranded RNA (dsRNA) effectively induces RNAi in insects, comprehensive studies on how small interfering RNA (siRNA) structural characteristics influence messenger RNA (mRNA) cleavage efficiency are limited. This study systematically examined the impact of diverse sequence and structural modifications on the efficiency of siRNA-mediated knockdown of target genes in Drosophila melanogaster. Our findings indicate that siRNA efficacy drastically decreased at a length of 17 nucleotides (nt), but was restored by extending to 19 base pairs (bp) with random sequences. Additionally, siRNAs with 2-nt overhangs demonstrated greater efficacy compared to blunt-ended structures. We found that the knockdown efficiency varies depending on the secondary structure characteristics of the mRNA region to which siRNA binds. Furthermore, next-generation sequencing was employed to map predicted siRNA distributions to actual dsRNA processing patterns, allowing detailed profiling of cleavage depth and sequence preferences. Collectively, these results enhance our understanding of the relationship between siRNA sequence design parameters and RNAi efficiency in Drosophila, providing a significant advance in siRNA-based RNAi research in insects. This study also offers valuable insights into dsRNA off-target effects.

大量的研究已经阐明了昆虫体内RNA干扰（RNAi）的机制。尽管双链RNA （dsRNA）能有效诱导昆虫体内的RNAi，但关于小干扰RNA （siRNA）结构特征如何影响信使RNA （mRNA）切割效率的全面研究仍然有限。本研究系统地研究了不同序列和结构修饰对黑腹果蝇siRNA敲除效率的影响。我们的研究结果表明，siRNA的功效在长度为17个核苷酸（nt）时显著下降，但通过随机序列扩展到19个碱基对（bp）后恢复。此外，与钝端结构相比，具有2-nt悬垂的sirna表现出更大的功效。相应的mRNA位点在sirna中特异性结构，降低了敲除效率。此外，下一代测序被用于将预测的siRNA分布映射到实际的dsRNA加工模式，从而可以详细分析切割深度和序列偏好。总的来说，这些结果增强了我们对果蝇siRNA序列设计参数与RNAi效率之间关系的理解，为基于siRNA的昆虫RNAi研究提供了显著的优势。这项研究也为dsRNA脱靶效应提供了有价值的见解。

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引用次数: 0

Apis mellifera multi-omics responses to ectoparasitic mites: unveiling biomarkers for Tropilaelaps mercedesae and Varroa destructor in honeybee hemolymph through hemolymph peptide and proteome analysis using MALDI-MS and LC-ESI-MS/MS 应用MALDI-MS和LC-ESI-MS/MS对蜜蜂血淋巴中Tropilaelaps mercedesae和Varroa destructor进行肽和蛋白质组学分析，揭示蜜蜂血淋巴中Tropilaelaps和Varroa destructor的生物标志物。

IF 3.7 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2025-10-09 DOI: 10.1016/j.ibmb.2025.104416

Patcharin Phokasem , Michel Bocquet , Sébastien N. Voisin , Chainarong Sinpoo , Camille Houdelet , Panuwan Chantawannakul , Terd Disayathanoowat , Philippe Bulet

Varroa destructor and Tropilaelaps mercedesae are ectoparasites that pose a significant threat to honeybee populations. It is essential to understand their impact to develop effective management strategies. In this study, we examined the relative abundance of proteins and the expression of immune-related genes in A. mellifera parasitized by the two mites using mass spectrometry analysis of hemolymph and transcriptomics of bleeding honeybees. The study revealed the modulation of genes and proteins associated with immune response and stress management. The apidaecin gene was upregulated in Varroa-infested honeybees and downregulated in Tropilaelaps-infested honeybees, a finding that correlated with MALDI-MS data. Furthermore, variations in the abundance of Hexamerin, Odorant Binding Protein 21, Phenoloxidase and Transferrin were observed in Varroa- and Tropilaelaps-infested honeybees. Using proteomics, potential markers associated with immunity, metabolism and stress responses (e.g. high Glx storage protein, alpha-glucosidase, glyoxylate/hydroxypyruvate reductases) were found to be specific to Varroa and Tropilaelaps infestations. Two Deformed Wing Virus (DWV) proteins exhibit higher abundance in the hemolymph of Tropilaelaps-infested honeybees compared to non-infested and Varroa-infested honeybees, suggesting that Tropilaelaps may weaken honeybee immunity, potentially exacerbating viral infections and colony health decline. This indicates that studying host-mite species interactions could lead to a better understanding of honeybee protection.

破坏瓦螨和粘虫是对蜜蜂种群构成严重威胁的寄生昆虫。了解它们对制定有效管理战略的影响是至关重要的。在本研究中，我们利用血淋巴质谱分析和出血蜜蜂转录组学检测了被这两种螨虫寄生的蜜蜂中蛋白质的相对丰度和免疫相关基因的表达。该研究揭示了与免疫反应和压力管理相关的基因和蛋白质的调节。蜂素基因在被瓦螨侵染的蜜蜂中表达上调，在被tropilaelaps侵染的蜜蜂中表达下调，这一发现与MALDI-MS数据相关。此外，在被瓦螨和tropilaelapers侵染的蜜蜂中，观察到六聚氰胺、气味结合蛋白21、酚氧化酶和转铁蛋白丰度的变化。利用蛋白质组学，发现了与免疫、代谢和应激反应相关的潜在标记（如高Glx储存蛋白、α -葡萄糖苷酶、乙醛酸盐/羟丙酮酸还原酶）对Varroa和Tropilaelaps侵袭具有特异性。与未感染和瓦罗亚感染的蜜蜂相比，被Tropilaelaps感染的蜜蜂的血淋巴中有两种变形翼病毒（DWV）蛋白的丰度更高，这表明Tropilaelaps可能会削弱蜜蜂的免疫力，潜在地加剧病毒感染和群体健康下降。这表明，研究宿主与螨虫物种的相互作用可以更好地理解蜜蜂的保护。

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引用次数: 0

Kinetic studies of pheromone-binding proteins (LdisPBP1 and LdisPBP2) from spongy moth Lymantria dispar with natural ligands and their analogues: Enantiomer distinction at external and internal binding sites Lymantria dispar海绵蛾信息素结合蛋白（LdisPBP1和LdisPBP2）与天然配体及其类似物的动力学研究：内外结合位点的对映体区分

IF 3.7 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2025-10-09 DOI: 10.1016/j.ibmb.2025.104419

Erika Plettner, Ashna Aulakh , Mailyn Terrado

Olfaction plays an important role in the reproduction of the spongy moth, Lymantria dispar, a major defoliator of deciduous trees. Detection of the attractant sex pheromone, (7R, 8S)-epoxy-2-methyloctadecane, (+)-disparlure, aids males in the location of mates by triggering upwind flight behavior. The antipode, (−)-disparlure, inhibits upwind flight behavior. The sensory hairs on the antennae of the male moth are innervated by the dendrite of an olfactory neuron, which is immersed in sensory lymph, a solution with pheromone-binding proteins (PBPs). These proteins bind pheromone molecules selectively and are necessary for proper olfactory functioning. Two PBPs from spongy moth are known: LdisPBP1 and LdisPBP2. The two proteins exhibit differences in their binding affinities towards various natural and unnatural ligands at their internal binding sites. Ligands are bound by PBPs in a two-step process: ligand capture results in binding to an external site, which is followed by movement of the ligand to the internal site. In this study, we investigate the differences in association and dissociation rates of the two proteins with (+)-disparlure, its enantiomer, (−)-disparlure, and some analogues with heteroatom substitutions of oxygen or sulfur at either carbon-5 or carbon-10, using stopped-flow kinetics. We used a fluorescent reporter (N-phenyl-1-naphthylamine, NPN), which competes with the ligands of interest and reports on external site binding in PBPs. The results are discussed in the context of kinetic selectivity of PBPs towards natural disparlure and synthetic analogue ligands, at their external binding sites, where enantiomer distinction has not been detected previously. Ligand docking simulations at two external sites of A and B forms and at one internal site of the B forms of the two proteins were used to interpret the enantioselectivity in the association and dissociation of the ligands. This paper also discusses the enantiomer selectivity and the molecular role of PBPs in the desorption from hydrophobic surfaces and partition between lipid vesicles and aqueous solutions of the pheromone enantiomers and their heteroatom-substituted analogues. Results are interpreted in terms of the contribution of ligand association and dissociation kinetics to the enantiomer selectivity of the spongy moth PBPs.

嗅觉在海绵蛾（Lymantria dispar）的繁殖中起着重要作用，Lymantria dispar是落叶树的主要落叶者。检测引诱性信息素(7R, 8S)-环氧-2-甲基十八烷，(+)-异引诱，通过引发逆风飞行行为来帮助雄性定位配偶。对跖点，(-)-差异，抑制逆风飞行行为。雄蛾触角上的感觉毛由嗅觉神经元的树突支配，该神经元浸在感觉淋巴中，这是一种含有信息素结合蛋白（pbp）的溶液。这些蛋白质选择性地结合信息素分子，是正常嗅觉功能所必需的。海绵蛾中已知两种PBPs: LdisPBP1和LdisPBP2。这两种蛋白质在其内部结合位点对各种天然和非天然配体的结合亲和力表现出差异。配体与PBPs结合的过程分为两步：配体捕获导致与外部位点结合，然后配体移动到内部位点。在这项研究中，我们使用停止流动动力学研究了两种蛋白质与(+)-disparlure，它的对映体，(-)-disparlure和一些类似物在碳-5或碳-10上杂原子取代氧或硫的结合和解离率的差异。我们使用了一种荧光报告因子（n -苯基-1-萘胺，NPN），它与感兴趣的配体竞争，并报告了PBPs中外部位点的结合。这些结果是在PBPs对天然差异和合成类似配体的动力学选择性的背景下讨论的，在它们的外部结合位点上，对映体的区别以前没有被检测到。在A和B两种蛋白的两个外部位点和B两种蛋白的一个内部位点进行配体对接模拟，以解释配体在结合和解离中的对映选择性。本文还讨论了对映体的选择性以及PBPs在信息素对映体及其异原子取代类似物的疏水表面解吸和脂质囊泡与水溶液之间的分割中的分子作用。根据配体结合和解离动力学对海绵蛾PBPs对映体选择性的贡献，对结果进行了解释。

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引用次数: 0

Female accessory reproductive glands of Paederus fuscipes serve as a reservoir of symbiotic pederin-producing bacteria 雌性副生殖腺作为共生的产童精细菌的储存库。

IF 3.7 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2025-10-02 DOI: 10.1016/j.ibmb.2025.104408

Xuhao Song , Hui Meng , Tingbang Yang , Yujie Li , Fake Zheng , Xianghui Yan

Paederus fuscipes, an ecologically and medically important species, is known for its blistering toxin pederin in hemolymph. Evidence demonstrates that the toxin is synthesized by the uncultured symbiotic pederin-producing bacteria (PPB) in P. fuscipes, but the biological characteristics of PPB within the beetle host remain poorly characterized. Here, we investigated PPB abundance variations in P. fuscipes across different factors (sexes, life stages, habitats, and organs), along with their colonization sites and metabolic potentials. The findings revealed that the PPB abundance in female P. fuscipes at the level of individuals and tissues exhibited stable colonization patterns, independent of habitat and time changes. Notably, PPB dominated the bacterial community in females (relative abundance ≥66.08 %) and nearly occupied reproductive organs (relative abundance ≥96.31 %). Moreover, our results indicated that PPB were predominantly enriched in the accessory glands of female reproductive organs, which could serve as a reservoir for PPB proliferation. Although PPB were not cultured in this study, metagenomic binning yielded the draft genome of PPB (CheckM completeness = 85.14 %, contamination = 0), in which genes related to pederin biosynthesis were identified. Phylogenetic analyses revealed that PPB formed a sister clade to Pseudomonas aeruginosa rather than nesting within the P. aeruginosa lineage. Metabolic module prediction analysis revealed specific deficiencies in PPB's energy metabolism and amino acid biosynthesis pathways, suggesting limited free-living potential for PPB. Collectively, this study provides insights into PPB biological characteristics within their beetle host and paves the way for biotechnological exploitation related to pederin production.

fuscipes Paederus是一种重要的生态和医学物种，以其在血淋巴中的起泡毒素pederin而闻名。有证据表明，该毒素是由fuscipes中未培养的共生pederin产生细菌（PPB）合成的，但PPB在甲虫宿主中的生物学特性仍然不清楚。在这里，我们研究了不同因素（性别、生命阶段、栖息地和器官）以及它们的定植地点和代谢潜力之间PPB丰度的变化。结果表明，在个体水平和组织水平上，雌性云杉的PPB丰度呈现稳定的定殖模式，不受生境和时间变化的影响。值得注意的是，PPB菌群在雌性中占主导地位（相对丰度≥66.08%），几乎占据生殖器官（相对丰度≥96.31%）。此外，我们的研究结果表明，PPB主要富集于女性生殖器官的附属腺，这可能是PPB增殖的储存库。虽然本研究没有培养PPB，但宏基因组测序得到了PPB的草图基因组（CheckM完整性= 85.14%，污染= 0），其中鉴定出了与pederin生物合成相关的基因。系统发育分析表明，PPB形成了铜绿假单胞菌的姐妹分支，而不是在铜绿假单胞菌谱系中筑巢。代谢模块预测分析揭示了PPB能量代谢和氨基酸生物合成途径的特异性缺陷，表明PPB的自由生活潜力有限。总的来说，本研究提供了PPB在其甲虫宿主内的生物学特性的见解，并为与pederin生产相关的生物技术开发铺平了道路。

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引用次数: 0

Determining the type of aerobic/anaerobic metabolism during exercise in Drosophila using incremental loading exercise 使用增量负荷运动确定果蝇运动期间的有氧/无氧代谢类型。

IF 3.7 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2025-10-01 DOI: 10.1016/j.ibmb.2025.104404

Qin Yi , Meng Ding , Jinglin Liu, Xiaoya Wang, Lan Zheng

Currently, Drosophila exercise is only classified as regular or endurance exercise, and there is no aerobic or anaerobic form of exercise, making Drosophila exercise modeling studies limited. Therefore, this study aimed to determine the type of aerobic and anaerobic exercise in Drosophila using Incremental Load exercise. A seven-day-old wild-type Drosophila was used as a study sample for exercise intervention using a Drosophila exercise device.

This study was designed with six exercise intensities, including 0.31 rev/s (E1), 0.45 rev/s (E2), 0.59 rev/s (E3), 0.71 rev/s (E4), 0.83 rev/s (E5), 0.91 rev/s (E6), and Control in addition Drosophila in all exercise groups performed a one-time 2.5-h acute exercise, and the control group did no exercise intervention. Anaerobic and aerobic metabolic enzymes in Drosophila were assayed immediately after exercise. Additionally, Drosophila exercise types were determined using hemolymph lactate, respiratory quotient, trehalose concentration, and mitochondrial respiration assays.

The experimental results show that at E2 exercise intensity, Drosophila mainly uses aerobic metabolism, and at E6 exercise intensity, it mainly uses anaerobic metabolism.We exercised Drosophila for 2 weeks at E2 and E6 exercise intensities to examine whether the physiological differences (cardiac function, climbing ability, sleep ability, lifespan, etc.) were consistent with those observed in mammals undergoing long-term aerobic and anaerobic exercise.The results showed that these differences are consistent with mammals.

目前，果蝇运动仅被归类为常规或耐力运动，并没有有氧或无氧运动形式，这使得果蝇运动建模研究受到限制。因此，本研究旨在通过增量负荷运动来确定果蝇的有氧和无氧运动类型。一只7天大的野生型果蝇作为研究样本，使用果蝇运动装置进行运动干预。本研究设计了0.31 rev/s （E1）、0.45 rev/s （E2）、0.59 rev/s （E3）、0.71 rev/s （E4）、0.83 rev/s （E5）、0.91 rev/s （E6）和对照组6种运动强度。此外，所有运动组果蝇进行一次2.5 h急性运动，对照组不进行运动干预。运动后立即测定果蝇的无氧和有氧代谢酶。此外，使用血淋巴乳酸、呼吸商、海藻糖浓度和线粒体呼吸测定来确定果蝇的运动类型。实验结果表明，在E2运动强度下，果蝇主要利用有氧代谢，在E6运动强度下，果蝇主要利用无氧代谢。我们以E2和E6运动强度对果蝇进行了为期两周的锻炼，以检验其生理差异（心功能、攀爬能力、睡眠能力、寿命等）是否与长期进行有氧和无氧运动的哺乳动物一致。结果表明，这些差异与哺乳动物一致。

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引用次数: 0

The role of the transformer gene in sex determination and its employment in CRISPR/Cas9-based homing gene drive in the global fruit pest Drosophila suzukii 变形基因在全球水果害虫铃木果蝇性别决定中的作用及其在CRISPR/ cas9基因驱动中的应用

IF 3.7 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2025-09-25 DOI: 10.1016/j.ibmb.2025.104406

Dan Deng , Xueying Yi , Wen Wen, Liuqing He, Wei Peng

Sex determination of Diptera is established by the cascade genes such as transformer (tra), though the primary signals for sex determination differ among different insects. Here, we report the isolation, expression and function of tra gene in an invasive pest, Drosophila suzukii, and study the potential use of the D. suzukii tra (Dstra) gene in CRISPR/Cas9-based homing gene drive for genetic-based pest management. The Dstra gene is highly conserved in structure and has a sex-specific transcript. To test the function of this gene in sex determination, Dstra dsRNA was injected into embryos. Almost all XX embryos developed into masculinized phenotypic male adults with intersex morphology. Abnormal ovaries were revealed in XX pseudomales upon dissection. Based on the necessary role of Dstra for female development, we developed and evaluated a homing gene drive that targets Dstra in D. suzukii. The drive component consisting of multiplex Dstra single guide RNAs and Cas9 with Dsvasa promoter was introduced into the Dstra locus. Abnormal development of both the external genitalia and gonads was observed in G0 and G1 chromosomal female adults that expressed the male-specific doublesex (dsx) transcript. Interestingly, knocking out Dstra led to significantly reduced fertility in adults of corresponding sex and moderate transmission rates of the DsRed gene (63.54 %) were observed. Our results not only confirm the conserved function of the Dstra gene in sex determination, but also highlight the potential of sex conversion-based suppression gene-drive strategy targeting the Dstra gene in controlling of D. suzukii populations.

双翅目昆虫的性别决定是由变压器（tra）等级联基因决定的，但不同昆虫性别决定的主要信号不同。本文报道了入侵害虫铃木果蝇tra基因的分离、表达和功能，并研究了铃木果蝇tra （Dstra）基因在基于CRISPR/ cas9的归巢基因驱动中用于害虫遗传管理的潜在应用。Dstra基因在结构上高度保守，并具有性别特异性转录。为了测试该基因在性别决定中的功能，将Dstra dsRNA注入胚胎。几乎所有的XX胚胎发育为阳刚化表型的雄性成虫，具有双性形态。XX例假男性经解剖发现卵巢异常。基于Dstra在女性发育中的必要作用，我们开发并评估了一种针对Dstra的铃木氏线虫归巢基因驱动。将多重Dstra单导rna和带有Dsvasa启动子的Cas9组成的驱动组件引入Dstra基因座。在G0和G1染色体上表达雄性特异性双性（dsx）转录本的成年雌性中，外生殖器和性腺发育异常。有趣的是，敲除Dstra导致相应性别的成人生育能力显著降低，并且观察到DsRed基因的中等传播率（63.54%）。我们的研究结果不仅证实了Dstra基因在性别决定中的保守功能，而且强调了以Dstra基因为目标的基于性别转换的抑制基因驱动策略在控制铃木夜蛾种群中的潜力。

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引用次数: 0