Insect Biochemistry and Molecular Biology最新文献_第7页

Biotin deficiency bridges metabolic signaling to insecticide sequestration in Nilaparvata lugens 生物素缺乏架起了褐飞虱体内代谢信号与杀虫剂隔离的桥梁。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2026-01-01 Epub Date: 2025-11-08 DOI: 10.1016/j.ibmb.2025.104445

Mengqing Deng, Xiyue Xu, Zhiming Yang, Kai Lu

Insecticide resistance poses a critical challenge to global agricultural sustainability. While metabolic detoxification and target-site mutations are well-characterized resistance mechanisms, the role of micronutrient homeostasis remains understudied. This study reveals that biotin deficiency in Nilaparvata lugens drives imidacloprid resistance through a multi-tiered regulatory network coordinating chemosensory protein (CSP) dynamics. Biotin deficiency enhances CSP-mediated insecticide sequestration via high-affinity binding to CSP2, CSP4, CSP7, and CSP15, which are overexpressed in resistant strains. RNA interference and dual-luciferase assays demonstrate that the aryl hydrocarbon receptor and its nuclear translocator (AhR/ARNT) transcriptionally activate CSP2 and CSP15, with their knockdown partially restoring insecticide susceptibility. Furthermore, biotin deficiency activates reactive oxygen species (ROS)/cap ‘n’ collar C (CncC) signaling, elevating AhR/ARNT expression through transcriptional reprogramming. Yeast three-hybrid assays identify a post-translational regulatory layer, wherein biotin directly inhibits AhR–ARNT heterodimerization. Field-evolved resistant populations recapitulate this mechanism, exhibiting conserved molecular signatures including biotin deficiency, ROS/CncC pathway activation, and AhR/ARNT-CSP overexpression correlated with resistance intensity. These findings establish a unified model wherein biotin scarcity reprograms xenobiotic defense through three synergistic mechanisms: enhanced CSP–insecticide binding, transcriptional amplification via ROS/CncC-AhR/ARNT signaling, and post-translational optimization of transcriptional complexes. The operational conservation of this pathway across laboratory and ecological contexts underscores its evolutionary significance while revealing novel targets for resistance management, particularly biotin-based synergists and AhR dimerization inhibitors.

杀虫剂抗药性对全球农业可持续性构成重大挑战。虽然代谢解毒和靶点突变是抗性机制的特征，但微量营养素稳态的作用仍未得到充分研究。本研究揭示了虫螺生物素缺乏通过协调化学感觉蛋白（CSP）动态的多层调控网络驱动吡虫啉抗性。生物素缺乏通过与抗性菌株中过表达的CSP2、CSP4、CSP7和CSP15的高亲和力结合，增强了csp介导的杀虫剂隔离。RNA干扰和双荧光素酶实验表明，芳烃受体及其核转运子（AhR/ARNT）转录激活CSP2和CSP15，其敲除部分恢复了对杀虫剂的敏感性。此外，生物素缺乏激活活性氧(ROS)/cap 'n' collar C （CncC）信号，通过转录重编程提高AhR/ARNT的表达。酵母三杂交实验鉴定了翻译后调控层，其中生物素直接抑制AhR-ARNT异源二聚化。田间进化的抗性群体概括了这一机制，表现出保守的分子特征，包括生物素缺乏、ROS/CncC途径激活以及与抗性强度相关的AhR/ARNT-CSP过表达。这些发现建立了一个统一的模型，其中生物素稀缺性通过三种协同机制重新编程外源防御：增强csp -杀虫剂结合，通过ROS/ cc - ahr /ARNT信号进行转录扩增，转录复合物的翻译后优化。该途径在实验室和生态环境中的操作保护强调了其进化意义，同时揭示了耐药性管理的新靶点，特别是基于生物素的增效剂和AhR二聚化抑制剂。

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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 : 2025-12-01 Epub 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

V-ATPase B mediates Bt Cry1Ac binding and toxicity in Grapholita molesta V-ATPase B介导Bt - Cry1Ac的结合和毒性作用。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-12-01 Epub Date: 2025-09-24 DOI: 10.1016/j.ibmb.2025.104407

Dandan Pan , Jiayang Feng , Jiacheng Ye, Xiaoyan Yang, Xiaoyan Zhang, Xiaofang Sha, Donghan Wang, Yanshen Fu, Chunxiao Men, Xiangqun Yuan, Yiping Li

Grapholita molesta is a worldwide pest. Cry1Ac is a significant alternative Bacillus thuringiensis (Bt) protein that exhibits substantial toxicity towards Lepidoptera insects. This investigation employed ligand blot, SDS-PAGE and liquid chromatography-tandem mass spectrometry (LC-MS/MS) techniques to identify the proteins potentially binding Cry1Ac on the peritrophic membrane (PM) of G. molesta. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis revealed the highest expression of GmolV-ATPase B in the midgut of 4th instar larvae. Following the ingestion of Cry1Ac protoxin by G. molesta larvae, a notable reduction in the expression level of GmolV-ATPase B was observed. The interaction between GmolV-ATPase B and activated Cry1Ac toxin was confirmed through ligand blot and homologous and heterologous competition experiments. Overexpression of GmolV-ATPase B in Sf9 cells led to an increase in Cry1Ac cytotoxicity, while RNAi targeting GmolV-ATPase B resulted in reduced mortality. In vivo bioassays demonstrated that the combined action of GmolV-ATPase B protein and Cry1Ac protoxin significantly enhanced the toxicity of Cry1Ac towards G. molesta larvae compared to Cry1Ac alone. These findings shed light on the binding of Cry1Ac to PM of G. molesta and its insecticidal mechanism, offering a valuable important reference for the development of biopesticides targeting midgut PM proteins.

墨氏笔蝗是一种世界性的害虫。Cry1Ac是一种重要的苏云金芽孢杆菌（Bacillus thuringiensis, Bt）替代蛋白，对鳞翅目昆虫具有显著的毒性。本研究采用配体印迹、SDS-PAGE和液相色谱-串联质谱（LC-MS/MS）技术，鉴定了鼠梨周围营养膜（PM）上可能结合Cry1Ac的蛋白。实时定量聚合酶链反应（RT-qPCR）结果显示，GmolV-ATPase B在4龄幼虫中肠的表达量最高。结果表明，食入Cry1Ac原毒素后，GmolV-ATPase B的表达量显著降低。GmolV-ATPase B与活化的Cry1Ac毒素通过配体印迹和同源、异源竞争实验证实了相互作用。Sf9细胞中过表达GmolV-ATPase B导致Cry1Ac细胞毒性增加，而靶向GmolV-ATPase B的RNAi导致死亡率降低。体内生物实验表明，与单独使用Cry1Ac相比，gmolv - atp酶B蛋白与Cry1Ac原蛋白的联合作用显著增强了Cry1Ac对鼠夜蛾幼虫的毒性。这些研究结果揭示了Cry1Ac与molesta的PM结合及其杀虫机制，为开发靶向中肠PM蛋白的生物农药提供了有价值的重要参考。

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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 : 2025-12-01 Epub 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

HSF-1 regulates FoxO expression to induce diapause in the cotton bollworm, Helicoverpa armigera via upstream HIF-1α and MNK signaling hif -1通过上游HIF-1α和MNK信号调控棉铃虫FoxO表达诱导滞育。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-12-01 Epub Date: 2025-10-27 DOI: 10.1016/j.ibmb.2025.104429

Zhi-Ren Su, Xiao-Long Su, Song-Shan Jiang, Wei-Hua Xu

Diapause is a widespread adaptation of insects that allows for them to survive under environmental stress by activating different gene clusters to inhibit metabolism and delay development. Previous studies have shown that reactive oxygen species (ROS) in brains of diapause-destined pupae of the cotton bollworm Helicoverpa armigera activate hypoxia-inducible factor-1α (HIF-1α), a transcription factor, to induce diapause. However, the molecular mechanism by which HIF-1α induces diapause remains unclear. In this study, a high abundance of heat shock factor-1 (HSF-1) protein, a transcription factor, was detected in brains of diapause-destined pupae, and phosphorylated HSF-1 was its active form. HSF-1 was phosphorylated by the protein kinase MNK, a member of the mitogen-activated protein kinase (MAPK) family. Moreover, the transcription of MNK was regulated by HIF-1α. Activated HSF-1 then promoted the expression of FoxO, which is the master regulator that triggers slow development to induce diapause. These results suggest that the HIF-1α/MNK/HSF-1/FoxO regulatory pathway promotes insect diapause.

滞育是昆虫的一种广泛的适应性，通过激活不同的基因簇来抑制新陈代谢和延迟发育，使它们能够在环境胁迫下生存。已有研究表明，棉铃虫滞育蛹大脑中的活性氧（ROS）可激活转录因子缺氧诱导因子-1α (HIF-1α)诱导滞育。然而，HIF-1α诱导滞育的分子机制尚不清楚。本研究在滞育蛹的大脑中检测到高丰度的转录因子热休克因子-1 （HSF-1）蛋白，磷酸化的HSF-1是其活性形式。HSF-1被蛋白激酶MNK磷酸化，MNK是丝裂原活化蛋白激酶（MAPK）家族的一员。此外，MNK的转录受HIF-1α的调控。激活的HSF-1随后促进FoxO的表达，FoxO是触发缓慢发育诱导滞育的主要调控因子。这些结果表明HIF-1α/MNK/HSF-1/FoxO调控通路促进昆虫滞育。

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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 : 2025-12-01 Epub 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

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 : 2025-12-01 Epub 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

Screening and evaluation of promoters for effective expression of fluorescent protein and high performance of the modified hyPBase in Zeugodacus cucurbitae 葫芦鱼荧光蛋白高效表达启动子筛选与评价及改性hyPBase的高效表达。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-12-01 Epub Date: 2025-10-22 DOI: 10.1016/j.ibmb.2025.104425

Siya Ma , Lixin Wang , Ying Yang , Zhongshi Zhou , Zizhen Fan , Yan Wu , Fengqin Cao , Xianwu Lin , Rihui Yan

The melon fly (Zeugodacus cucurbitae(Coquillett)) is a highly destructive quarantine pest that inflicts significant damage on the agricultural industry. However, the absence of efficient genetic research tools for this pest hinders its research progress and therefore its control. Among these, the selection of an appropriate promoter plays a central role. In this study, the activities of numerous promoter systems were examined by transfecting Drosophila S2 cells and microinjecting Z. cucurbitae embryos. The results reveal that native promoters drive higher expression within their respective species than in others and that the ZcPUb promoter effectively drives the expression of fluorescent proteins in Z. cucurbitae. A hyperactive piggyBac transposase (hyPBase) was also genetically modified to the control of the ZcPUb promoter and it significantly improved the transgenesis efficiency for constructs of different sizes. The temporal and spatial expression patterns of fluorescence regulated by the ZcPUb promoter in transgenic flies suggest that the ZcPUb promoter-controlled fluorescence can be used as a valuable marker for transgenic screening of Z. cucurbitae. These findings not only highlight the ZcPUb promoter as a crucial resource for future investigations in gene functions and developmental processes in Z. cucurbitae, but also emphasize the importance of identifying native promoters for research and practical application of organisms.

瓜蝇（Zeugodacus cucurbitae(Coquillett)）是一种高度破坏性的检疫性害虫，对农业造成重大损害。然而，缺乏有效的遗传研究工具阻碍了这种害虫的研究进展，从而阻碍了其控制。其中，选择合适的启动子起着核心作用。在本研究中，通过转染果蝇S2细胞和微注射葫芦瓜卵，检测了多个启动子系统的活性。结果表明，原生启动子在各自物种中的表达高于其他启动子，ZcPUb启动子有效地驱动了Z. cucurbitae荧光蛋白的表达。我们还对一种高度活跃的piggyBac转座酶（hyPBase）进行了基因修饰，以控制ZcPUb启动子，并显著提高了不同大小构建体的转基因效率。ZcPUb启动子调控的荧光在转基因蝇中的时空表达模式表明，ZcPUb启动子调控的荧光可作为筛选葫芦虱转基因的一种有价值的标记。这些发现不仅突出了ZcPUb启动子作为未来研究葫芦瓜基因功能和发育过程的重要资源，也强调了鉴定原生启动子对生物研究和实际应用的重要性。

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引用次数: 0

Genome-wide analysis of the spider Pardosa pseudoannulata revealed the function of spliceosome components in cold adaptation 对假环蛛的全基因组分析揭示了剪接体成分在冷适应中的功能。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-12-01 Epub Date: 2025-10-22 DOI: 10.1016/j.ibmb.2025.104426

Jingjing Li , Na Yu , Yunru Chen , Zewen Liu

Organisms adapt to cold stress in winter with various molecular mechanisms. Alternative splicing (AS) is one of the most extensive mechanisms in response to cold stress in plants, but has been rarely reported in animals. Arthropods are widely distributed on earth, and overwintering poses a challenge for the population growth. Here, we investigated the role of AS in response to cold stress in arthropods using the spider Pardosa pseudoannulata as a model. We found that about 48.7 % of multi-exon genes underwent AS events in overwintering spiders collected from fields, and ∼35.3 % were alternatively spliced in cold-acclimated spiders in laboratory, showing a gradual increase as temperature decreases. Spliceosome components were concurrently regulated by cold stress in both transcription and AS aspects. Knockdown of SMB/B′ and SMFa during cold acclimation impaired the regulation on AS of cold-responsive genes and led to reduced cold stress tolerance. SMB/B’ and SMFa improved cold adaptation probably through regulating AS events of genes from 4 enriched pathways (spliceosome, circadian rhythm, circadian rhythm-fly, and RNA degradation), which have been reported important in tolerance to cold stress. Taken together, spliceosome components contribute to cold acclimation by ensuring the adequate splicing patterns of cold-responsive genes in the spider, and enhanced its tolerance to cold stress.

生物适应冬季冷胁迫的分子机制多种多样。选择性剪接（Alternative splicing， AS）是植物对冷胁迫反应最广泛的机制之一，但在动物中很少报道。节肢动物在地球上分布广泛，越冬对其种群增长构成了挑战。本文以假环蜘蛛为研究对象，研究了AS在节肢动物冷应激反应中的作用。我们发现，在野外采集的越冬蜘蛛中，约48.7%的多外显子基因发生了AS事件，而在实验室中，约35.3%的多外显子基因发生了选择性剪接，并随着温度的降低而逐渐增加。剪接体组分在转录和AS方面同时受到冷胁迫的调控。在冷驯化过程中，SMB/B'和SMFa基因的表达下调会影响冷响应基因对AS的调节，导致冷胁迫耐受性降低。SMB/B'和SMFa可能通过调节4个富集途径（剪接体、昼夜节律、昼夜节律蝇和RNA降解）基因的AS事件来改善冷适应，这些途径在耐受冷胁迫中很重要。综上所述，剪接体成分通过确保蜘蛛中冷响应基因的适当剪接模式来促进冷适应，并增强其对冷胁迫的耐受性。

{"title":"Genome-wide analysis of the spider Pardosa pseudoannulata revealed the function of spliceosome components in cold adaptation","authors":"Jingjing Li , Na Yu , Yunru Chen , Zewen Liu","doi":"10.1016/j.ibmb.2025.104426","DOIUrl":"10.1016/j.ibmb.2025.104426","url":null,"abstract":"<div><div>Organisms adapt to cold stress in winter with various molecular mechanisms. Alternative splicing (AS) is one of the most extensive mechanisms in response to cold stress in plants, but has been rarely reported in animals. Arthropods are widely distributed on earth, and overwintering poses a challenge for the population growth. Here, we investigated the role of AS in response to cold stress in arthropods using the spider <em>Pardosa pseudoannulata</em> as a model. We found that about 48.7 % of multi-exon genes underwent AS events in overwintering spiders collected from fields, and ∼35.3 % were alternatively spliced in cold-acclimated spiders in laboratory, showing a gradual increase as temperature decreases. Spliceosome components were concurrently regulated by cold stress in both transcription and AS aspects. Knockdown of <em>SMB/B′</em> and <em>SMFa</em> during cold acclimation impaired the regulation on AS of cold-responsive genes and led to reduced cold stress tolerance. SMB/B’ and SMFa improved cold adaptation probably through regulating AS events of genes from 4 enriched pathways (spliceosome, circadian rhythm, circadian rhythm-fly, and RNA degradation), which have been reported important in tolerance to cold stress. Taken together, spliceosome components contribute to cold acclimation by ensuring the adequate splicing patterns of cold-responsive genes in the spider, and enhanced its tolerance to cold stress.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"185 ","pages":"Article 104426"},"PeriodicalIF":3.7,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145367237","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}

引用次数: 0

A single Drosophila Dscam1 genomic locus generates a vast repertoire of circRNAs facilitated by RNA pairing 单个果蝇Dscam1基因组位点通过RNA配对产生大量环状RNA。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-12-01 Epub Date: 2025-10-23 DOI: 10.1016/j.ibmb.2025.104423

Zedong Li , Bingbing Xu , Zhechao Wang , Wenjie Zhang , Jiayan Fu , Lili Wu , Nengcheng Bao , Jinpeng Xie , Ru Yan , Haiyang Dong , Yongfeng Jin

Drosophila melanogaster Dscam1 gene has the potential to generate 38,016 linear isoforms through mutually exclusive alternative splicing. Here, using a tailored approach, we identified thousands of circRNA isoforms derived from the Dscam1 locus, representing the highest number of circRNA isoforms produced by a single gene to date. The extensive organization of alternative circularization of Dscam1s is conserved within Insecta. Notably, the analysis of circRNA isoforms revealed that alternative back-splicing of variable exon 4s and 6s occurs in a proximity-dependent manner. Interestingly, unlike the classical “loop-in” model mediated by RNA pairing between flanking introns, we revealed a “loop-out” mechanism, in which RNA secondary structures between docking sites and selector sequences facilitate the back-splicing of exons located outside the loop. These findings expand the understanding of the mechanism underlying the generation of circRNA and provide new insights into the expression regulation of highly complex gene loci.

果蝇Dscam1基因有可能通过互斥的选择性剪接产生38,016个线性同种异构体。在这里，使用定制的方法，我们鉴定了来自Dscam1位点的数千种circRNA异构体，代表了迄今为止单个基因产生的最多数量的circRNA异构体。在昆虫纲中，Dscam1s的交替循环的广泛组织是保守的。值得注意的是，对circRNA异构体的分析显示可变外显子4s和6s的选择性反向剪接以邻近依赖的方式发生。有趣的是，与经典的由侧翼内含子之间的RNA配对介导的“环入”模型不同，我们揭示了一种“环出”机制，其中对接位点和选择序列之间的RNA二级结构促进了环外外显子的反向剪接。这些发现扩大了对circRNA产生机制的理解，并为高度复杂基因位点的表达调控提供了新的见解。

{"title":"A single Drosophila Dscam1 genomic locus generates a vast repertoire of circRNAs facilitated by RNA pairing","authors":"Zedong Li , Bingbing Xu , Zhechao Wang , Wenjie Zhang , Jiayan Fu , Lili Wu , Nengcheng Bao , Jinpeng Xie , Ru Yan , Haiyang Dong , Yongfeng Jin","doi":"10.1016/j.ibmb.2025.104423","DOIUrl":"10.1016/j.ibmb.2025.104423","url":null,"abstract":"<div><div><em>Drosophila melanogaster Dscam1</em> gene has the potential to generate 38,016 linear isoforms through mutually exclusive alternative splicing. Here, using a tailored approach, we identified thousands of circRNA isoforms derived from the <em>Dscam1</em> locus, representing the highest number of circRNA isoforms produced by a single gene to date. The extensive organization of alternative circularization of <em>Dscam1</em>s is conserved within <em>Insecta</em>. Notably, the analysis of circRNA isoforms revealed that alternative back-splicing of variable exon 4s and 6s occurs in a proximity-dependent manner. Interestingly, unlike the classical “loop-in” model mediated by RNA pairing between flanking introns, we revealed a “loop-out” mechanism, in which RNA secondary structures between docking sites and selector sequences facilitate the back-splicing of exons located outside the loop. These findings expand the understanding of the mechanism underlying the generation of circRNA and provide new insights into the expression regulation of highly complex gene loci.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"185 ","pages":"Article 104423"},"PeriodicalIF":3.7,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145370080","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}

引用次数: 0