Insect Biochemistry and Molecular Biology最新文献_第2页

Starvation induces a transition from autophagy to apoptosis via the ER-Ca2+-calpain signaling axis in the fat body of Bombyx mori 饥饿通过家蚕脂肪体中的ER-Ca2+-calpain信号轴诱导自噬向凋亡的转变

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

Insect Biochemistry and Molecular Biology

Pub Date : 2026-01-12 DOI: 10.1016/j.ibmb.2026.104494

Jialu Cheng , Sheng Zhang , Haoyi Gu , Xueling Qin , Bing Li

Nutritional stress is a common environmental challenge for insects in nature. As a central organ of energy metabolism, the fat body plays a crucial role in maintaining energy homeostasis. However, the mechanisms by which the fat body regulates programmed cell death (PCD) during energy crises remain unclear. In this study, the fat body of Bombyx mori was used as a model to investigate the regulatory mechanisms underlying PCD transitions induced by sustained energy depletion. The results indicated that starvation caused a significant reduction in adenosine triphosphate (ATP), rapid depletion of glycogen and triglycerides, and marked inhibition of the endoplasmic reticulum (ER) calcium pump enzyme SERCA. Concurrently, it upregulated the expression of the calcium channel IP3R. These combined effects led to Ca²⁺ efflux from the ER calcium stores and cytoplasmic Ca²⁺ overload. Intracellular Ca²⁺ levels significantly increased, followed by a decrease during prolonged starvation, paralleling changes in calpain activity and accompanied by the upregulation of the proapoptotic protein NtATG5. Short-term starvation promoted autophagy by significantly increasing LC3-II and ATG5 expression, whereas prolonged starvation promoted calpain-mediated cleavage of ATG5 to generate NtATG5 and activated caspase-3, leading to apoptosis. Furthermore, the IP3R inhibitor 2-APB significantly suppressed starvation-induced calcium signaling, autophagy, and apoptosis. This study reveals the regulatory mechanism by which the Bombyx mori fat body modulates PCD under starvation stress via the ER-Ca²⁺-calpain signaling pathway, providing crucial insights into how nutritional deprivation causes physiological damage in insects.

营养压力是自然界昆虫普遍面临的环境挑战。脂肪体作为能量代谢的中枢器官，在维持能量稳态中起着至关重要的作用。然而，脂肪体在能量危机期间调节程序性细胞死亡（PCD）的机制尚不清楚。本研究以家蚕的脂肪体为模型，探讨持续能量消耗诱导PCD转变的调控机制。结果表明，饥饿导致三磷酸腺苷（ATP）显著降低，糖原和甘油三酯迅速消耗，内质网钙泵酶SERCA明显抑制。同时上调钙离子通道IP3R的表达。这些综合作用导致内质网钙储存的Ca2+外排和细胞质Ca2+超载。细胞内Ca2+水平显著升高，随后在长时间饥饿期间下降，与钙蛋白酶活性的变化平行，并伴有促凋亡蛋白NtATG5的上调。短期饥饿通过显著增加LC3-II和ATG5的表达来促进自噬，而长期饥饿通过calpain介导的ATG5裂解生成NtATG5并激活caspase-3导致细胞凋亡。此外，IP3R抑制剂2-APB显著抑制饥饿诱导的钙信号、自噬和细胞凋亡。本研究揭示了家蚕脂肪体在饥饿胁迫下通过ER-Ca2+-calpain信号通路调节PCD的调控机制，为了解营养剥夺如何导致昆虫的生理损伤提供了重要的见解。

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引用次数: 0

NPF-OBP5 regulate sugar sensitivity and feeding behavior of rice planthoppers NPF-OBP5调控稻飞虱的糖敏感性和摄食行为

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

Insect Biochemistry and Molecular Biology

Pub Date : 2026-01-12 DOI: 10.1016/j.ibmb.2026.104495

Ruichuan Duan, Yewei Jia, Xinru Xiao, Shuqi Ren, Guijun Wan, Fajun Chen

Neuropeptide F (NPF) is considered to be related to the feeding behavior and odorant binding protein (OBP) is very important for the chemical perception of herbivorous insects to host plants, while, food sugar is a strong inducer for insect feeding. However, it is unknown whether NPF and OBP can regulate the sugar perception, thus affecting the feeding behavior and sugar metabolism in brown planthopper (BPH) Nilaparvata lugens. The results showed that NlOBP1, NlOBP5-8 and NlOBP10 were highly expressed in N. lugens heads. Fluorescence competitive binding assays demonstrated that only NlOBP5 exhibited strong combination to soluble sugars (including glucose, fructose, and sucrose). Additionally, NlNPF was predominantly expressed in N. lugens brain. NlNPF knockdown resulted in upregulated NlOBP5 expression and enhanced the sensitivity of N. lugens to soluble sugars. Conversely, NlOBP5 knockdown reduced the sugar sensitivity of N. lugens, while simultaneous knockdown of both NlNPF and NlOBP5 partially rescued the sugar sensitivity impairment caused by NlOBP5 knockdown individually. Furthermore, NlNPF knockdown promoted feeding and led to trehalose accumulation in N. lugens. In contrast, NlOBP5 knockdown inhibited feeding and decreased trehalose levels in N. lugens, while simultaneous knockdown of NlNPF and NlOBP5 partially reversed the negative effects on feeding and trehalose metabolism observed with NlOBP5 knockdown individually. NlNPF knockdown decreased the survival of N. lugens more than NlOBP5 knockdown. These findings confirm the regulation of NlNPF-NlOBP5 on sugar sensitivity, feeding behavior and trehalose metabolism of N. lugens, providing potential targets for the control strategies on rice planthoppers.

神经肽F （NPF）被认为与取食行为有关，气味结合蛋白（OBP）在草食性昆虫对寄主植物的化学感知中起着重要作用，而食糖是昆虫取食的强诱导剂。然而，NPF和OBP是否能调节褐飞虱的糖感知，从而影响褐飞虱的摄食行为和糖代谢尚不清楚。结果表明，NlOBP1、NlOBP5-8和NlOBP10在褐飞虱头中高表达。荧光竞争结合实验表明，只有NlOBP5与可溶性糖（包括葡萄糖、果糖和蔗糖）有很强的结合。此外，NlNPF主要表达于N. lugens脑中。NlNPF敲低导致NlOBP5表达上调，增强了N. lugens对可溶性糖的敏感性。相反，NlOBP5敲低降低了N. lugens的糖敏感性，而NlNPF和NlOBP5同时敲低部分挽救了NlOBP5单独敲低引起的糖敏感性损伤。NlNPF基因敲低促进了褐藻糖的摄食，导致褐藻糖在褐藻菌体内积累。相比之下，NlOBP5敲低抑制了N. lugens的摄食并降低了海藻糖水平，而NlNPF和NlOBP5同时敲低部分逆转了NlOBP5单独敲低观察到的对摄食和海藻糖代谢的负面影响。NlNPF敲低比NlOBP5敲低更能降低N. lugens的存活率。这些发现证实了NlNPF-NlOBP5对褐飞虱糖敏感性、摄食行为和海藻糖代谢的调控作用，为水稻飞虱防治策略提供了潜在靶点。

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引用次数: 0

Trachealess, a bHLH-PAS transcription factor, is an upstream regulator of 20-hydroxyecdysone signaling essential for molting in Henosepilachna vigintioctopunctata Trachealess是一种bHLH-PAS转录因子，是20-羟基蜕皮激素信号传导的上游调节因子，对Henosepilachna viintioctopunctata的蜕皮至关重要。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2026-01-10 DOI: 10.1016/j.ibmb.2026.104493

Junna Liu , Qi Chen , June-Sun Yoon , Mujuan Guo , Chunxiao Yang , Xuguo Zhou , Youjun Zhang , Huipeng Pan

The steroid hormone 20-hydroxyecdysone (20E) is the central regulator of insect molting, yet the upstream transcription factors controlling its pathway remain poorly characterized. Here, we investigate the role of the bHLH-PAS transcription factor Trachealess (HvTrh) in the 28-spotted ladybeetle, Henosepilachna vigintioctopunctata. HvTrh was ubiquitously expressed during larval development, with peak transcript levels in the pupal stage. RNAi-mediated silencing of HvTrh induced severe molting defects and high mortality across all larval instars. This was associated with a significant reduction in 20E titers and the suppression of key 20E pathway genes. The resulting molting defects were not rescued by exogenous 20E, indicating a disruption of tissue competence. Histological analysis confirmed that HvTrh knockdown impaired chitin deposition and new cuticle remodeling. Among the downregulated genes, HvE75 was the most strongly suppressed. Mechanistically, we demonstrated that HvTrh directly activates the HvE75 promoter by binding to a specific central nervous system midline element (CME), as validated by dual-luciferase reporter assays and electrophoretic mobility shift assays. Our findings establish HvTrh as a critical upstream regulator of insect molting, proposing a model where it governs the 20E signaling cascade, at least in part, through the direct transcriptional control of HvE75.

类固醇激素20-羟基蜕皮激素（20E）是昆虫蜕皮的中心调节因子，但控制其途径的上游转录因子仍不清楚。在此，我们研究了bHLH-PAS转录因子Trachealess （HvTrh）在28斑瓢虫Henosepilachna vigintioctopunctata中的作用。HvTrh在幼虫发育过程中普遍表达，在蛹阶段转录水平达到峰值。rnai介导的HvTrh沉默在所有幼虫中引起严重的蜕皮缺陷和高死亡率。这与20E滴度的显著降低和关键20E途径基因的抑制有关。由此产生的蜕皮缺陷不能通过外源性20E修复，表明组织能力受到破坏。组织学分析证实，HvTrh敲低会损害几丁质沉积和新角质层重塑。在下调基因中，HvE75受到的抑制最为强烈。在机制上，我们证明了HvTrh通过结合特定的中枢神经系统中线元件（CME）直接激活HvE75启动子，这一点通过双荧光素酶报告基因试验和电泳迁移转移试验得到了验证。我们的研究结果证实HvTrh是昆虫蜕化的关键上游调控因子，并提出了一个模型，其中HvTrh至少部分通过直接转录控制HvE75来调控20E信号级联。

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引用次数: 0

DIL-CRISPR: a practical approach to mitigate G0 mosaic lethality in insect gene editing DIL-CRISPR：减轻昆虫基因编辑中G0花叶致死率的实用方法

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

Insect Biochemistry and Molecular Biology

Pub Date : 2026-01-09 DOI: 10.1016/j.ibmb.2026.104492

Hiiragi Nishizawa, Takaaki Daimon

Genome editing in insects is typically conducted by injecting genome editing reagents into early embryos, producing generation zero (G₀) individuals that develop as genetic mosaics. Targeting genes whose disruption induces mosaic lethality is therefore challenging, since most or all G₀ individuals frequently fail to survive to adulthood, preventing germline transmission of edited alleles. Here, we present a straightforward and practical approach, DIL-CRISPR, to mitigate G₀ mosaic lethality by systematically diluting the CRISPR/Cas9 injection mix. Using the tobacco cutworm Spodoptera litura and the juvenile hormone receptor gene Met1 as a benchmark, we demonstrate that dilution of the injection mix lessens the severity of larval–pupal mosaic phenotypes and increases G₀ survival in a dose-dependent manner. Amplicon sequencing further showed that somatic mutation frequencies decline with dilution, while germline mutation rates remain sufficient to establish mutant lines. Notably, we detected a substantial discrepancy between somatic and germline editing efficiencies, likely reflecting selective loss of highly edited, lethal mosaics before they reach adulthood. We conclude that DIL-CRISPR therefore offers a reliable means to balance G₀ survival with germline editing, converting an empirically used dilution practice into a generalizable strategy. Overall, this approach provides a practical solution for generating mutant lines of mosaic lethal genes and is broadly applicable across diverse insect species, facilitating functional genetic studies in non-model insects.

昆虫的基因组编辑通常是通过将基因组编辑试剂注射到早期胚胎中，产生作为遗传嵌合体发育的第0代（G0）个体来进行的。因此，靶向破坏可诱导花叶致死的基因具有挑战性，因为大多数或所有G0个体经常无法存活到成年，从而阻止了编辑等位基因的种系传播。在这里，我们提出了一种简单实用的方法，DIL-CRISPR，通过系统地稀释CRISPR/Cas9注射混合物来减轻G0嵌合致死率。以斜纹夜蛾（Spodoptera litura）和幼虫激素受体基因Met1为基准，我们证明了稀释后的注射混合物降低了幼虫-蛹马赛克表型的严重程度，并以剂量依赖的方式增加了G0存活率。扩增子测序进一步表明，体细胞突变频率随着稀释而下降，而种系突变率仍然足以建立突变系。值得注意的是，我们发现体细胞和种系编辑效率之间存在巨大差异，这可能反映了高度编辑的致命性嵌合体在成年之前的选择性损失。因此，我们得出结论，DIL-CRISPR提供了一种可靠的方法来平衡G0存活和种系编辑，将经验上使用的稀释实践转化为一种可推广的策略。总之，该方法为产生花叶致死基因突变系提供了一种实用的解决方案，并且广泛适用于不同昆虫物种，促进了非模式昆虫的功能遗传研究。

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引用次数: 0

Collagen type XV regulation via MMP2-dependent degradation governs ovarian development in the rice stem borer, Chilo suppressalis XV型胶原蛋白通过mmp2依赖性降解调控水稻螟虫卵巢发育。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2026-01-08 DOI: 10.1016/j.ibmb.2026.104490

Yue Li , Suijie Kuang , Yan Tang , Zhuotian Yao , Yechen Pan , Ziren Zang , Yang Cui , Hongshuai Gao , Lin Qiu , Yansong Xiao , Tin Liu , Qi Su , Youzhi Li

Female reproductive development is fundamental to the remarkable reproductive capacity of insects, and extracellular matrix (ECM) remodeling plays a critical role in ovarian development. However, the function of Collagen, a major ECM component, in female insect ovarian development remains poorly characterized. Using the major rice pest Chilo suppressalis as a model, we investigated the role of Col XV in insect reproduction. We cloned and characterized Cscol15, a gene encoding a 1077-amino acid protein. Real-time quantitative PCR analysis revealed peak Cscol15 expression during the egg stage and was significantly higher in the head of female pupae compared to other examined tissues. RNAi-mediated knockdown of the Cscol15 gene in female C. suppressalis pupae severely disrupted ovarian development, manifesting as reduced ovarian length and width, decreased vitellogenin deposition, and lower collagen fiber abundance. To further validate these findings, we employed CRISPR/Cas9-mediated knockout of Cscol15, which confirmed that its loss impairs ovarian development and significantly compromises female fertility in C. suppressalis.As a key regulator of collagen synthesis and degradation, we further investigated the role of MMP2 in regulating Collagen XV. Our analysis revealed that overexpression of CsMMP2 reduced the level of Collagen XV, thereby impairing insect reproductive capacity. These findings establish Collagen XV as a key ECM component, likely directly degraded by MMP2. This mechanism governs ovarian morphogenesis in C. suppressalis, positioning Collagen XV as a promising target for reproductive disruption in pest management strategies.

雌性生殖发育是昆虫卓越生殖能力的基础，而细胞外基质（ECM）重塑在卵巢发育中起着至关重要的作用。然而，胶原蛋白（ECM的主要成分）在雌性昆虫卵巢发育中的作用仍然不清楚。以水稻主要害虫抑制螟（Chilo suppressalis）为模型，研究了colxv在昆虫繁殖中的作用。我们克隆并鉴定了一个编码1077个氨基酸的蛋白的基因ccol15。实时定量PCR分析显示，ccol15的表达在卵期达到峰值，且在雌蛹头部的表达量明显高于其他检测组织。rnai介导的ccol15基因敲低严重影响卵巢发育，表现为卵巢长度和宽度减小，卵黄原蛋白沉积减少，胶原纤维丰度降低。为了进一步验证这些发现，我们利用CRISPR/ cas9介导的基因敲除ccol15，证实了ccol15的缺失会损害卵巢发育，并显著影响C. suppressalis的雌性生育能力。作为胶原合成和降解的关键调节因子，我们进一步研究了MMP2在调节XV胶原中的作用。我们的分析表明，CsMMP2的过表达降低了XV胶原蛋白的水平，从而损害了昆虫的生殖能力。这些发现证实胶原XV是关键的ECM成分，可能直接被MMP2降解。这种机制控制着C. suppressalis的卵巢形态发生，将胶原XV定位为害虫管理策略中生殖破坏的有希望的目标。

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引用次数: 0

TostOBP12 mediates the olfactory response to host volatiles in the parasitoid wasp Trichogramma ostriniae TostOBP12介导拟寄生蜂赤眼蜂对寄主挥发物的嗅觉反应

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

Insect Biochemistry and Molecular Biology

Pub Date : 2026-01-07 DOI: 10.1016/j.ibmb.2026.104491

Xiang-xin Kong , Hou-xuan Zhang , Hong-yu Liang , Han Xiao , Yu-qin Wang , Shuang Gang , Hui Dong

Trichogramma wasps are widely utilized as critical biological control agents in agricultural systems. Its efficacy relies on detecting host-derived chemical cues, yet the key molecular determinants underlying this process remain elusive. Here, we demonstrate that the odorant-binding protein TostOBP12 is indispensable for the host-seeking behavior of T. ostriniae. Interspecific behavioral assays revealed that only T. ostriniae exhibited significantly enhanced parasitism driven by the host sex pheromone. TostOBP12 was identified as the most highly expressed OBP in female heads. Molecular docking and competitive binding assays confirmed its high and broad affinity for both sex pheromones ((Z)-12:14AC and (E)-12:14AC) and egg surface compounds. Crucially, RNAi-mediated silencing of TostOBP12 completely abolished the wasp's behavioral response to sex pheromones without affecting its development, confirming its essential in vivo function. Our findings reveal a key molecular sensor in a tritrophic interaction and provide a promising target for enhancing biocontrol efficacy.

赤眼蜂作为重要的生物防治剂在农业系统中得到了广泛的应用。它的功效依赖于检测宿主衍生的化学线索，但这一过程背后的关键分子决定因素仍然难以捉摸。在这里，我们证明了气味结合蛋白TostOBP12在ostriniae的寻宿主行为中是不可或缺的。种间行为分析表明，在寄主性信息素的驱动下，只有玉米螟蛾表现出明显的寄生性增强。TostOBP12是女性头部中表达量最高的OBP。分子对接和竞争结合实验证实了其对性信息素(（Z)-12:14AC和(E)-12:14AC）和卵子表面化合物的高亲和力和广泛亲和力。至关重要的是，rnai介导的TostOBP12沉默完全消除了黄蜂对性信息素的行为反应，而不影响其发育，证实了其在体内的基本功能。我们的发现揭示了一个在营养相互作用中的关键分子传感器，并为提高生物防治效果提供了一个有希望的靶点。

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引用次数: 0

Trichlorfon induces damage of growth and development of Bombyx mori through ferroptosis pathway 敌百虫通过死亡途径对家蚕生长发育造成损害。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2026-01-05 DOI: 10.1016/j.ibmb.2026.104488

Wen Zhang , Huang Lin , Ziqin Xiao , Dongxu Shen , Dingguo Xia , Zhiyong Qiu , Xuelian Zhang , Qiaoling Zhao

Trichlorfon (TCF), an organophosphorus pesticide, has long been used in agriculture as an insecticide for crop protection. However, its residual presence has caused significant economic losses to the sericulture industry, hindering the development of Bombyx mori (B. mori) farming. Although previous studies have investigated the toxicity of TCF to silkworms, the detailed toxic effects and molecular mechanisms remain poorly understood. In this study, the potential molecular targets and mechanisms of TCF-induced injury in silkworms were investigated in vivo and in vitro by combining RNA sequencing, qRT-PCR and other techniques. It is found that TCF exposure leads to damage of growth and development in silkworms, as evidenced by reduced body weight, increased mortality, and decline of cluster, cocooning, pupation and egg production. Further RNA sequencing analysis identified several differentially expressed genes, particularly LOC101744260, as well as altered pathways, notably the glutamine metabolism pathway—both of which are closely associated with ferroptosis. In vivo and in vitro experiments have confirmed that the expression levels of ferroptosis-related genes, including Fer HCH, Fer 2LCH, Keap1, Tf, Gtpx and SOD1 exhibited significant changes in expression levels. These changes are accompanied by elevated intracellular Fe²⁺ and Fe³⁺ levels and oxidative stress. Moreover, N-acetylcysteine (NAC) treatment partially reverses TCF-induced growth and developmental impairments at the individual and cellular levels. Taken together, these results indicate for the first time that TCF exposure induces damage of growth and development in silkworms by activating the ferroptosis pathway. This study provides a new insight into the toxic mechanisms of TCF in silkworms and offers a theoretical basis for the prevention and control of pesticide pollution in silkworms industry.

敌百虫（TCF）是一种有机磷农药，长期以来在农业中作为作物保护杀虫剂使用。然而，它的残留给桑蚕业造成了重大的经济损失，阻碍了家蚕养殖业的发展。虽然以前的研究已经调查了TCF对家蚕的毒性，但详细的毒性作用和分子机制仍然知之甚少。本研究结合RNA测序、qRT-PCR等技术，在体内和体外研究了tcf诱导家蚕损伤的潜在分子靶点和机制。研究发现，接触TCF会损害家蚕的生长发育，表现为体重下降、死亡率升高、成团、结茧、化蛹和产蛋量下降。进一步的RNA测序分析发现了几个差异表达的基因，特别是LOC101744260，以及改变的途径，特别是谷氨酰胺代谢途径，两者都与铁下垂密切相关。体内和体外实验证实，ferhch、fer2lch、Keap1、Tf、Gtpx、SOD1等铁中毒相关基因的表达水平发生了显著变化。这些变化伴随着细胞内Fe2+和Fe3+水平升高和氧化应激。此外，n -乙酰半胱氨酸（NAC）治疗在个体和细胞水平上部分逆转了tcf诱导的生长和发育障碍。综上所述，这些结果首次表明TCF暴露通过激活铁下垂途径诱导家蚕生长发育损伤。本研究对TCF对家蚕的毒性机制提供了新的认识，并为家蚕行业农药污染的防治提供了理论依据。

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引用次数: 0

Functional characterization of voltage-gated sodium channels in two mirid pests and identification of kdr mutations in pyrethroid resistant Lygus lineolaris population 电压门控钠通道在两种杂交害虫中的功能特征及拟除虫菊酯抗性线螟种群中kdr突变的鉴定。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2026-01-05 DOI: 10.1016/j.ibmb.2026.104486

Yuzhe Du , Shane Scheibener , Yu-Cheng Zhu , Omaththage P. Perera , Gadi V.P. Reddy , Boris S. Zhorov

Lygus lineolaris and Lygus hesperus are economically important pests that cause substantial damage to wide variety of crops in the United States. In this study, we isolated 17 full-length voltage-gated sodium channels cDNA clones encoding 14 unique splicing variants from a susceptible strain of L. hesperus (sLhNav), two variants from a susceptible L. lineolaris strain (sLlNav), and 8 clones encoding 7 variants from a pyrethroid-resistant L. lineolaris population (rLlNav), based on the usage of 12 alternative exons. Functional expression in Xenopus oocytes revealed that 11 sLh/LlNav variants produced sodium currents suitable for electrophysiological analysis, which displayed distinct gating properties. Notably, Nav variants lacking exon b generated significantly larger currents compared to those including exon b. We also identified A-to-I editing events in both Lygus Navs, and a key finding was the identification of three kdr mutations in rLlNav variants, L1023 F/S in segment IIS6 and L934I in the IIS4-S5 linker (corresponding to L1014F, L1014S and L925I in housefly Nav), all of which have been previously associated with pyrethroid resistance in other insect species. This is the first report of these mutations in resistant L. lineolaris populations. Together with earlier reports of enhanced metabolic detoxification, our findings suggest a dual resistance strategy in L. lineolaris, involving both target-site insensitivity and elevated detoxification enzyme activity. The comprehensive characterization of sodium channel variants in both Lygus, along with the identification of L1023 F/S and L934I mutations, provides valuable insight into Nav diversity in Lygus species and the molecular basis of pyrethroid resistance in L. lineolaris.

在美国，线尾灰盲蝽和大尾灰盲蝽是经济上重要的害虫，对多种作物造成严重损害。在这项研究中，我们基于12个替代外显子的使用，分离了17个全长电压门控钠通道cDNA克隆，编码14个独特的剪接变体，来自L. hesperus易感菌株（sLhNav），来自L. lineolaris易感菌株（sLlNav）的2个变体，以及来自L. lineolaris抗性群体（rLlNav）的8个克隆，编码7个变体。在爪蟾卵母细胞中的功能表达表明，11种sLh/LlNav变体产生了适合电生理分析的钠电流，表现出不同的门控特性。值得注意的是，缺乏外显子b的Nav变异比含有外显子b的Nav变异产生的电流大得多。我们还在两种Lygus Nav中发现了a -to- i编辑事件，一个关键发现是在rLlNav变异中发现了三个kdr突变，即IIS6片段的L1023F/S和IIS4-S5连接子的L934I（对应于家蝇Nav中的L1014F、L1014S和L925I），所有这些突变之前都与其他昆虫物种的拟除虫虫酯抗性有关。这是在耐药的线状乳杆菌群体中首次报道这些突变。结合早期的代谢解毒增强的报道，我们的发现表明lineolaris存在双重抗性策略，包括靶点不敏感和解毒酶活性升高。对这两种莱格斯钠通道变异的全面表征，以及L1023F/S和L934I突变的鉴定，为了解莱格斯物种中钠通道多样性和莱格斯拟除虫菊酯抗性的分子基础提供了有价值的见解。

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引用次数: 0

The cytochrome P450s CYP6CY3 and CYP6CY4 confer resistance to flupyradifurone in the green peach aphid Myzus persicae 细胞色素p450 CYP6CY3和CYP6CY4赋予桃蚜对氟吡喃酮的抗性。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2026-01-05 DOI: 10.1016/j.ibmb.2026.104487

Jun Yuan , Bin Zeng , Benjamin J. Hunt , Bartlomiej J. Troczka , Xingzhi Xiao , James Gardner , Faye Papadimitriou , Jason Charamis , Aris Ilias , Adam Pym , Konstantinos Mavridis , Dieudonné T. Tshitenge , Ralf Nauen , John Vontas , John T. Margaritopoulos , Xianhui Yin , Chris Bass

The green peach aphid, Myzus persicae, is an economically important crop pest that has evolved resistance to numerous insecticides used for control. Recent research has shown that populations of M. persicae in Greece have evolved resistance to flupyradifurone, a butenolide insecticide that acts as a reversible agonist on insect nicotinic acetylcholine receptors (nAChRs). However, the genetic mechanisms underpinning resistance remain unclear. Here we used transcriptomic profiling in combination with in vivo and in vitro functional approaches to show that flupyradifurone resistance in M. persicae is conferred by enhanced expression of the P450 genes CYP6CY3 and CYP6CY4. High levels of flupyradifurone resistance were observed in M. persicae clones from Greece and France, including in a clone that predates the registration of this insecticide. Synergist bioassays with piperonyl butoxide, an inhibitor of P450s, suggested resistance was mediated, in part, by enhanced P450 activity. Subsequent, transcriptome profiling identified several P450s that were overexpressed in the resistant M. persicae clones compared to susceptible clones. Functional characterisation of these P450s revealed that only CYP6CY3 and CYP6CY4 confer resistance in vivo and metabolise flupyradifurone in vitro. In two clones with extremely high levels of resistance to flupyradifurone, overexpression of these P450s was accompanied by the R81T mutation in the β1 subunit of the nAChR, known to confer resistance to neonicotinoids and cross-resistance to flupyradifurone. These findings provide new insight into the mechanisms of resistance to flupyradifurone and reveal how generalist P450s can be preadapted to confer cross-resistance to diverse insecticides.

桃蚜（Myzus persicae）是一种经济上重要的农作物害虫，它已经进化出了对许多用于控制的杀虫剂的抗性。最近的研究表明，希腊桃蚜种群已经进化出对氟吡喃醌的抗性，氟吡喃醌是一种丁烯内酯杀虫剂，可作为昆虫烟碱乙酰胆碱受体（nAChRs）的可逆激动剂。然而，支持抗性的遗传机制仍不清楚。在这里，我们使用转录组学分析结合体内和体外功能方法，表明桃蚜耐氟吡喃酮是通过P450基因CYP6CY3和CYP6CY4的表达增强而产生的。在希腊和法国的桃蚜无性系中观察到高水平的氟吡拉德酮抗性，包括在该杀虫剂登记之前的无性系。与胡椒酰丁醇（P450抑制剂）的协同生物测定表明，P450活性的增强部分介导了耐药性。随后，转录组分析发现，与易感克隆相比，抗性桃蚜克隆中有几个p450过表达。这些p450的功能特征表明，只有CYP6CY3和CYP6CY4在体内产生耐药性，并在体外代谢氟吡喃酮。在对氟吡地酮具有极高抗性的两个克隆中，这些p450的过表达伴随着nAChR β1亚基的R81T突变，已知该突变赋予对新烟碱类的抗性和对氟吡地酮的交叉抗性。这些发现提供了对氟吡喃酮抗性机制的新见解，并揭示了如何预先适应多面手p450以赋予对多种杀虫剂的交叉抗性。

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引用次数: 0

Structural and mechanistic insights into BoCSP1-mediated resistance to neonicotinoids in Bradysia odoriphaga 嗜臭迟缓虫bocsp1介导的新烟碱抗性的结构和机制研究

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

Insect Biochemistry and Molecular Biology

Pub Date : 2026-01-05 DOI: 10.1016/j.ibmb.2026.104489

Xingyu Ma, Junjie Zeng, Chunni Zhang, Wu Dai

The chemosensory protein (CSP)-mediated sequestration resistance mechanism has attracted increasing attention. This study revealed that a laboratory-selected clothianidin-resistant strain of Bradysia odoriphaga exhibited moderate cross-resistance to thiamethoxam. qPCR analysis demonstrated significant upregulation of BoCSP1 expression in the resistant strain, and its transcription could be induced by clothianidin exposure. RNAi-mediated silencing of BoCSP1 increased the susceptibility of resistant larvae to both clothianidin and thiamethoxam. Fluorescence competitive binding assays showed strong binding affinity between BoCSP1 and clothianidin/thiamethoxam, with Ki values < 3 μM. Molecular dynamics simulations and binding mode analysis further revealed that the stability of BoCSP1-clothianidin and BoCSP1-thiamethoxam complexes is primarily governed by van der Waals interactions. Computational alanine scanning (CAS) and site-directed mutagenesis identified Asn74 and Ile90 as critical residues mediating BoCSP1-clothianidin binding, while Trp101 was essential for BoCSP1-thiamethoxam interaction. Binding pocket analysis of the mutant proteins revealed that alanine substitutions disrupted hydrogen bonding networks and altered local conformational rigidity, leading to substantially reduced ligand affinity. These findings elucidate a novel CSPs-mediated resistance mechanism and provide a theoretical foundation for developing precision-engineered insecticides that exploit CSPs as unique molecular targets.

化学感觉蛋白（CSP）介导的固存抗性机制越来越受到人们的关注。本研究发现，实验室筛选的嗜臭迟缓虫耐噻虫胺菌株对噻虫胺表现出中度交叉抗性。qPCR分析显示，抗性菌株BoCSP1表达显著上调，其转录可能受到噻虫胺暴露的诱导。rnai介导的BoCSP1沉默增加了抗性幼虫对噻虫胺和噻虫嗪的敏感性。荧光竞争结合实验显示，BoCSP1与噻虫胺/噻虫胺具有较强的结合亲和力，Ki值< 3 μM。分子动力学模拟和结合模式分析进一步表明，bocsp1 -噻虫胺和bocsp1 -噻虫胺配合物的稳定性主要受范德华相互作用的控制。计算丙氨酸扫描（CAS）和定点突变鉴定出Asn74和Ile90是介导bocsp1 -噻虫胺结合的关键残基，而Trp101是bocsp1 -噻虫胺相互作用的必要残基。突变蛋白的结合袋分析显示，丙氨酸取代破坏了氢键网络，改变了局部构象刚性，导致配体亲和力大幅降低。这些发现阐明了一种新的CSPs介导的抗性机制，并为开发利用CSPs作为独特分子靶点的精准工程杀虫剂提供了理论基础。

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引用次数: 0