Insect Biochemistry and Molecular Biology最新文献_第9页

Circ-379-miR-3050-DNAH3/RNF8 axis modulates wing development in Aphis citricidus Circ-379-miR-3050-DNAH3/RNF8轴调控柑橘蚜虫翅膀发育

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-08-22 DOI: 10.1016/j.ibmb.2025.104393

Yu-Jing Liu , Jin-Ming Lu , Xiu-Cheng Xie , Qin-Qin Xu , Bi-Yue Ding , Jin-Jun Wang , Feng Shang

Wings are key physiological determinants of aphid dispersal and population outbreaks. Circular RNAs (circRNAs), a novel class of non-coding RNAs, regulate diverse biological processes through post-transcriptional mechanisms. However, their roles in insect wing development remain unclear. In this study, we performed RNA sequencing to identify 715 circRNAs across four developmental stages of Aphis citricidus: fourth-instar winged nymphs, winged adults, fourth-instar wingless nymphs, and wingless adults. Differential expression analysis revealed circ-379 as the only circRNA significantly upregulated during the fourth-instar winged nymphs to winged adults transition, while showing no significant change in wingless morphs, suggesting a potential role in wing development. Subsequent reverse transcription PCR (RT-PCR) validated the circular structure of circ-379 using divergent primers. Silencing of circ-379 induced wing deformities. Bioinformatic prediction, expression profiling analysis, dual-luciferase reporter assays and biotin-avidin-based RNA pull-down assays identified miR-3050 as a direct miRNA target of circ-379, which serves as a molecular sponge to bind miR-3050, thereby suppressing the expression of its target genes dynein axonemal heavy chain 3 (DNAH3) and E3 ubiquitin-protein ligase RNF8-B-like (RNF8). Overexpression of miR-3050 or RNA interference (RNAi)-mediated knockdown of DNAH3 and RNF8 also caused wing deformities. These findings indicate that circ-379 functions as a miR-3050 sponge to influence DNAH3 and RNF8 expression, thereby ensuring proper wing formation in A. citricidus. This study uncovers a circRNA-regulated molecular axis underlying wing plasticity in aphids, thereby advancing our understanding of non-coding RNA-mediated developmental regulation in insects.

翅是蚜虫扩散和种群爆发的关键生理决定因素。环状rna （circRNAs）是一类新型的非编码rna，通过转录后机制调节多种生物过程。然而，它们在昆虫翅膀发育中的作用尚不清楚。在这项研究中，我们对柑橘蚜虫的四个发育阶段（四龄有翅若虫、有翅成虫、四龄无翅若虫和无翅成虫）进行了RNA测序，鉴定了715个环状RNA。差异表达分析显示circ-379是唯一一个在四龄有翼若虫向有翼成虫转变过程中显著上调的circRNA，而在无翼形态中没有显著变化，这表明circ-379可能在翅膀发育中起作用。随后的反转录PCR （RT-PCR）使用发散引物验证了circ-379的圆形结构。circ-379诱导的机翼畸形的沉默。生物信息学预测、表达谱分析、双荧光素酶报告基因测定和基于生物素亲和素的RNA拉下测定鉴定miR-3050是circ-379的直接miRNA靶点，circ-379作为分子海绵结合miR-3050，从而抑制其靶基因动力蛋白轴索重链3 （DNAH3）和E3泛素蛋白连接酶RNF8- b样（RNF8）的表达。过表达miR-3050或RNA干扰（RNAi）介导的DNAH3和RNF8的敲低也会导致翅膀畸形。这些发现表明circ-379作为miR-3050海绵影响DNAH3和RNF8的表达，从而确保柑橘翅果的正确形成。本研究揭示了蚜虫翅膀可塑性背后的环状rna调控分子轴，从而促进了我们对非编码rna介导的昆虫发育调控的理解。

{"title":"Circ-379-miR-3050-DNAH3/RNF8 axis modulates wing development in Aphis citricidus","authors":"Yu-Jing Liu , Jin-Ming Lu , Xiu-Cheng Xie , Qin-Qin Xu , Bi-Yue Ding , Jin-Jun Wang , Feng Shang","doi":"10.1016/j.ibmb.2025.104393","DOIUrl":"10.1016/j.ibmb.2025.104393","url":null,"abstract":"<div><div>Wings are key physiological determinants of aphid dispersal and population outbreaks. Circular RNAs (circRNAs), a novel class of non-coding RNAs, regulate diverse biological processes through post-transcriptional mechanisms. However, their roles in insect wing development remain unclear. In this study, we performed RNA sequencing to identify 715 circRNAs across four developmental stages of <em>Aphis citricidus</em>: fourth-instar winged nymphs, winged adults, fourth-instar wingless nymphs, and wingless adults. Differential expression analysis revealed <em>circ-379</em> as the only circRNA significantly upregulated during the fourth-instar winged nymphs to winged adults transition, while showing no significant change in wingless morphs, suggesting a potential role in wing development. Subsequent reverse transcription PCR (RT-PCR) validated the circular structure of <em>circ-379</em> using divergent primers. Silencing of <em>circ-379</em> induced wing deformities. Bioinformatic prediction, expression profiling analysis, dual-luciferase reporter assays and biotin-avidin-based RNA pull-down assays identified miR-3050 as a direct miRNA target of <em>circ-379</em>, which serves as a molecular sponge to bind miR-3050, thereby suppressing the expression of its target genes <em>dynein axonemal heavy chain 3</em> (<em>DNAH3</em>) and <em>E3 ubiquitin-protein ligase RNF8-B-like</em> (<em>RNF8</em>). Overexpression of miR-3050 or RNA interference (RNAi)-mediated knockdown of <em>DNAH3</em> and <em>RNF8</em> also caused wing deformities. These findings indicate that <em>circ-379</em> functions as a miR-3050 sponge to influence <em>DNAH3</em> and <em>RNF8</em> expression, thereby ensuring proper wing formation in <em>A. citricidus</em>. This study uncovers a circRNA-regulated molecular axis underlying wing plasticity in aphids, thereby advancing our understanding of non-coding RNA-mediated developmental regulation in insects.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104393"},"PeriodicalIF":3.7,"publicationDate":"2025-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144907322","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

Study on the effects of high temperature on autophagy and apoptosis in different organs of Bombyx mori based on calcium ion homeostasis 基于钙离子稳态的高温对家蚕不同器官自噬和凋亡影响的研究

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-08-20 DOI: 10.1016/j.ibmb.2025.104384

Peilin Peng , Hongbin Zou , Jialu Cheng , Haoyi Gu , Guimin Nong , Xuelin Qin , Bing Li

High-temperature stress differentially affects physiological functions in Bombyx mori, including growth and development, silk protein synthesis, and metabolism. To investigate the underlying mechanisms, we explored the mechanisms of autophagy and apoptosis in the midgut, posterior silk gland (PSG), and fat body of B. mori under high-temperature treatment (30 °C and 35 °C), based on calcium ion (Ca²⁺) homeostasis. The results demonstrated that high-temperature treatment significantly reduced the body weight of B. mori. Furthermore, high-temperature treatment could cause damage to the midgut and PSG of B. mori, accompanied by intracellular Ca²⁺ overload, and the transcription levels of autophagy-related genes as well as the expression of LC3-I and LC3-II proteins are both affected. The protein levels of ATG5 and NtATG5 were significantly increased, while the transcription of key apoptotic genes and the protein levels of caspase-3 and cleaved caspase-3 were elevated. Collectively, these results indicated that high temperature induced autophagy in B. mori through Ca²⁺ mediation and promoted its conversion to apoptosis. Additionally, high temperature was demonstrated to inflict minimal damage on the fat body and failed to induce upregulation of autophagy or apoptosis. This study not only unveiled the intricate interplay among Ca²⁺ homeostasis, autophagy, and apoptosis in various organs of B. mori under high-temperature stress, but also revealed differential effects of heat stress across distinct organs. These findings provide a theoretical foundation for the breeding of thermotolerant silkworm varieties.

高温胁迫对家蚕生长发育、丝蛋白合成和代谢等生理功能有不同程度的影响。为了研究其潜在的机制，我们基于钙离子（Ca2+）稳态，研究了高温处理（30°C和35°C）下家蚕中肠、后丝腺（PSG）和脂肪体的自噬和凋亡机制。结果表明，高温处理显著降低了家蚕体重。此外，高温处理可导致家蚕中肠和PSG损伤，并伴有细胞内Ca2+超载，自噬相关基因的转录水平以及LC3-I和LC3-II蛋白的表达均受到影响。ATG5和NtATG5蛋白水平显著升高，凋亡关键基因转录及caspase-3和cleaved - caspase-3蛋白水平升高。综上所述，这些结果表明高温通过Ca2+介导诱导家蚕自噬，并促进其向凋亡的转化。此外，高温被证明对脂肪体造成的损伤最小，并且不能诱导自噬或凋亡的上调。本研究不仅揭示了高温胁迫下家蚕各器官Ca2+稳态、自噬和凋亡之间复杂的相互作用，而且揭示了热应激在不同器官间的差异效应。这些发现为选育耐热蚕品种提供了理论依据。

{"title":"Study on the effects of high temperature on autophagy and apoptosis in different organs of Bombyx mori based on calcium ion homeostasis","authors":"Peilin Peng , Hongbin Zou , Jialu Cheng , Haoyi Gu , Guimin Nong , Xuelin Qin , Bing Li","doi":"10.1016/j.ibmb.2025.104384","DOIUrl":"10.1016/j.ibmb.2025.104384","url":null,"abstract":"<div><div>High-temperature stress differentially affects physiological functions in <em>Bombyx mori</em>, including growth and development, silk protein synthesis, and metabolism. To investigate the underlying mechanisms, we explored the mechanisms of autophagy and apoptosis in the midgut, posterior silk gland (PSG), and fat body of <em>B. mori</em> under high-temperature treatment (30 °C and 35 °C), based on calcium ion (Ca<sup>2+</sup>) homeostasis. The results demonstrated that high-temperature treatment significantly reduced the body weight of <em>B. mori</em>. Furthermore, high-temperature treatment could cause damage to the midgut and PSG of <em>B. mori</em>, accompanied by intracellular Ca<sup>2+</sup> overload, and the transcription levels of autophagy-related genes as well as the expression of LC3-I and LC3-II proteins are both affected. The protein levels of ATG5 and NtATG5 were significantly increased, while the transcription of key apoptotic genes and the protein levels of caspase-3 and cleaved caspase-3 were elevated. Collectively, these results indicated that high temperature induced autophagy in <em>B. mori</em> through Ca<sup>2+</sup> mediation and promoted its conversion to apoptosis. Additionally, high temperature was demonstrated to inflict minimal damage on the fat body and failed to induce upregulation of autophagy or apoptosis. This study not only unveiled the intricate interplay among Ca<sup>2+</sup> homeostasis, autophagy, and apoptosis in various organs of <em>B. mori</em> under high-temperature stress, but also revealed differential effects of heat stress across distinct organs. These findings provide a theoretical foundation for the breeding of thermotolerant silkworm varieties.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104384"},"PeriodicalIF":3.7,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144889252","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

Transgressive gene expression disrupts the molting process in hybrids of two invasive termites 越界基因表达破坏了两种入侵白蚁杂交的蜕皮过程。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-08-15 DOI: 10.1016/j.ibmb.2025.104383

Kyung Seok Kim , Darshan Chetty , Thomas Chouvenc , Joseph F. Velenovsky IV , Sang-Bin Lee , Seema Rana , Edward L. Vargo

The Formosan subterranean termite (Coptotermes formosanus) and the Asian subterranean termite (Coptotermes gestroi) are among the most destructive termite pests in the world. Both species have spread to various regions worldwide with overlapping distributions in a few areas where they can potentially hybridize. Observations suggest that workers in hybrid colonies are slower to molt than those of the parental species, suggesting a disruption in the molting process as a form of hybrid incompatibility. Our goal was to identify misexpressed genes in hybrids during the molting process to help uncover the molecular mechanisms underlying molting disruption. We conducted RNA-seq and identified molting-related genes by performing a time course analysis on differentially expressed transcripts. We identified molting-related genes during each stage of the molting cycle (pre-, post- and inter-molt) in the parental species. We then compared expression levels of these genes in the hybrids to identify genes that were transgressively expressed (either over- or under-expressed) compared to the parental species. We identified several genes related to the molting cycle, muscle contraction, response to stress, and ecdysone metabolism that were under-expressed in hybrids relative to their parents. These differences may help explain the disruption of molting in hybrids and provide insights into the effects of hybridization on misexpression of genes during critical periods of growth and development. Moreover, identification of molting related genes in subterranean termites highlights the molecular pathways involved in the molting process in this group of insects with high developmental plasticity.

台湾地下白蚁（Coptotermes formosanus）和亚洲地下白蚁（Coptotermes gestroi）是世界上最具破坏性的白蚁害虫。这两个物种已经扩散到世界各地，在一些地区重叠分布，它们可以潜在地杂交。观察表明，杂交群体中的工蜂蜕皮速度比亲本物种慢，这表明蜕皮过程的中断是杂交不相容的一种形式。我们的目标是鉴定在蜕皮过程中杂种中错误表达的基因，以帮助揭示蜕皮中断的分子机制。我们进行了rna测序，并通过对差异表达转录物进行时间过程分析来鉴定蜕皮相关基因。我们在亲本物种的蜕皮周期的每个阶段（蜕皮前、蜕皮后和蜕皮间）鉴定了蜕皮相关基因。然后，我们比较了这些基因在杂交种中的表达水平，以确定与亲本物种相比，过度表达（或过表达或过表达）的基因。我们发现了几个与蜕皮周期、肌肉收缩、应激反应和蜕皮激素代谢相关的基因，这些基因在杂交后代中相对于亲代表达不足。这些差异可能有助于解释杂交种蜕皮的中断，并为杂交对生长发育关键时期基因错误表达的影响提供见解。此外，通过对地下白蚁蜕皮相关基因的鉴定，揭示了地下白蚁这种具有高度发育可塑性的昆虫蜕皮过程的分子途径。

{"title":"Transgressive gene expression disrupts the molting process in hybrids of two invasive termites","authors":"Kyung Seok Kim , Darshan Chetty , Thomas Chouvenc , Joseph F. Velenovsky IV , Sang-Bin Lee , Seema Rana , Edward L. Vargo","doi":"10.1016/j.ibmb.2025.104383","DOIUrl":"10.1016/j.ibmb.2025.104383","url":null,"abstract":"<div><div>The Formosan subterranean termite <em>(Coptotermes formosanus</em>) and the Asian subterranean termite (<em>Coptotermes gestroi</em>) are among the most destructive termite pests in the world. Both species have spread to various regions worldwide with overlapping distributions in a few areas where they can potentially hybridize. Observations suggest that workers in hybrid colonies are slower to molt than those of the parental species, suggesting a disruption in the molting process as a form of hybrid incompatibility. Our goal was to identify misexpressed genes in hybrids during the molting process to help uncover the molecular mechanisms underlying molting disruption. We conducted RNA-seq and identified molting-related genes by performing a time course analysis on differentially expressed transcripts. We identified molting-related genes during each stage of the molting cycle (pre-, post- and inter-molt) in the parental species. We then compared expression levels of these genes in the hybrids to identify genes that were transgressively expressed (either over- or under-expressed) compared to the parental species. We identified several genes related to the molting cycle, muscle contraction, response to stress, and ecdysone metabolism that were under-expressed in hybrids relative to their parents. These differences may help explain the disruption of molting in hybrids and provide insights into the effects of hybridization on misexpression of genes during critical periods of growth and development. Moreover, identification of molting related genes in subterranean termites highlights the molecular pathways involved in the molting process in this group of insects with high developmental plasticity.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"184 ","pages":"Article 104383"},"PeriodicalIF":3.7,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144861972","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

Evolution of a horizontally acquired fatty acid desaturase enables the biosynthesis of ω-3 polyunsaturated fatty acids in Collembola 水平获得的脂肪酸去饱和酶的进化使弹线虫能够生物合成ω-3多不饱和脂肪酸。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-08-07 DOI: 10.1016/j.ibmb.2025.104381

Yuya Ohhara , Ayumi Sato , Yoko Hirono-Hara , Kiyotaka Y. Hara , Kimiko Yamakawa-Kobayashi

Marine ecosystems are rich in essential dietary fatty acids, particularly ω-3 long-chain polyunsaturated fatty acids (PUFAs) including eicosapentaenoic acid (EPA). Various marine microorganisms including microalgae produce ω-3 long-chain PUFAs through the action of methyl- and front-end desaturases. However, most terrestrial organisms cannot produce ω-3 long-chain PUFAs due to the absence of methyl- and/or front-end desaturases. One notable exception is de novo EPA biosynthesis in springtails (Collembola), the non-insect soil hexapod lineage serving as a nutrient source for predatory animals; however, Collembola EPA biosynthesis enzymes remain unknown. Here, we provide the first evidence of the key desaturases required for EPA biosynthesis in Collembola. Mass spectrometric analysis suggested that EPA is synthesized through arachidonic acid biosynthesis and following ω-3 desaturation, which requires front- and methyl-end desaturases, respectively. RNA-sequencing of Collembola transcripts isolated multiple candidate genes encoding front-end desaturases; however, no clear orthologous sequence of a methyl-end desaturase was retrieved. Verification of the activity of the isolated enzymes using a yeast expression system revealed that Collembola front-end desaturases catalyzed arachidonic acid biosynthesis. Furthermore, a subgroup of Collembola front-end desaturase sequences catalyzed the ω-3 desaturation step, facilitating the bioconversion of arachidonic acid to EPA. Phylogenetic analysis further revealed that the Collembola front-end desaturase sequences clustered closely to those of marine microorganisms producing ω-3 long-chain PUFAs. Given that Collembola is derived from a marine Crustacea ancestor, we propose that the horizontal acquisition of a front-end desaturase gene from a marine microbe, followed by duplication and neofunctionalization, empowered Collembola to become an EPA producer.

海洋生态系统富含必需膳食脂肪酸，特别是ω-3长链多不饱和脂肪酸（PUFAs），其中包括二十碳五烯酸（EPA）。包括微藻在内的各种海洋微生物通过甲基和前端去饱和酶的作用产生ω-3长链PUFAs。然而，由于缺乏甲基和/或前端去饱和酶，大多数陆生生物不能产生ω-3长链pufa。一个值得注意的例外是在弹尾虫（弹尾虫）中重新合成EPA，这是一种非昆虫的土壤六足动物，是掠食性动物的营养来源；然而，线虫的EPA生物合成酶仍不为人所知。在这里，我们提供了线虫生物合成EPA所需的关键去饱和酶的第一个证据。质谱分析表明，EPA是通过花生四烯酸生物合成和ω-3脱饱和合成的，ω-3脱饱和分别需要前端和甲基端脱饱和酶。编码前端去饱和酶的多候选弹线虫转录本的rna测序然而，没有检索到甲基端去饱和酶的明确同源序列。利用酵母表达系统对分离酶的活性进行验证，发现弹线虫前端去饱和酶催化花生四烯酸的生物合成。此外，弹线虫前端去饱和酶序列的一个亚群催化ω-3去饱和步骤，促进花生四烯酸向EPA的生物转化。系统发育分析进一步表明，弹线虫前端去饱和酶序列与产生ω-3长链PUFAs的海洋微生物的去饱和酶序列密切相关。鉴于弹珠虫起源于海洋甲壳类动物祖先，我们认为从海洋微生物中水平获取前端去饱和酶基因，然后进行复制和新功能化，使弹珠虫成为EPA生产者。

{"title":"Evolution of a horizontally acquired fatty acid desaturase enables the biosynthesis of ω-3 polyunsaturated fatty acids in Collembola","authors":"Yuya Ohhara , Ayumi Sato , Yoko Hirono-Hara , Kiyotaka Y. Hara , Kimiko Yamakawa-Kobayashi","doi":"10.1016/j.ibmb.2025.104381","DOIUrl":"10.1016/j.ibmb.2025.104381","url":null,"abstract":"<div><div>Marine ecosystems are rich in essential dietary fatty acids, particularly ω-3 long-chain polyunsaturated fatty acids (PUFAs) including eicosapentaenoic acid (EPA). Various marine microorganisms including microalgae produce ω-3 long-chain PUFAs through the action of methyl- and front-end desaturases. However, most terrestrial organisms cannot produce ω-3 long-chain PUFAs due to the absence of methyl- and/or front-end desaturases. One notable exception is <em>de novo</em> EPA biosynthesis in springtails (Collembola), the non-insect soil hexapod lineage serving as a nutrient source for predatory animals; however, Collembola EPA biosynthesis enzymes remain unknown. Here, we provide the first evidence of the key desaturases required for EPA biosynthesis in Collembola. Mass spectrometric analysis suggested that EPA is synthesized through arachidonic acid biosynthesis and following ω-3 desaturation, which requires front- and methyl-end desaturases, respectively. RNA-sequencing of Collembola transcripts isolated multiple candidate genes encoding front-end desaturases; however, no clear orthologous sequence of a methyl-end desaturase was retrieved. Verification of the activity of the isolated enzymes using a yeast expression system revealed that Collembola front-end desaturases catalyzed arachidonic acid biosynthesis. Furthermore, a subgroup of Collembola front-end desaturase sequences catalyzed the ω-3 desaturation step, facilitating the bioconversion of arachidonic acid to EPA. Phylogenetic analysis further revealed that the Collembola front-end desaturase sequences clustered closely to those of marine microorganisms producing ω-3 long-chain PUFAs. Given that Collembola is derived from a marine Crustacea ancestor, we propose that the horizontal acquisition of a front-end desaturase gene from a marine microbe, followed by duplication and neofunctionalization, empowered Collembola to become an EPA producer.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"183 ","pages":"Article 104381"},"PeriodicalIF":3.7,"publicationDate":"2025-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144811506","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

The functional impact of mutations in orthosteric binding site of the Drosophila melanogaster Dα2-containing nicotinic receptors points to a greater contribution to neonicotinoid selectivity of an arginine in loop D of Dβ1 than an introduced serine in loop C of Dα2 黑腹果蝇含Dα2烟碱受体正位结合位点突变的功能影响表明，Dβ1环D中的精氨酸比Dα2环C中的引入丝氨酸对新烟碱选择性的贡献更大。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-08-06 DOI: 10.1016/j.ibmb.2025.104380

Ryo Ito , Hisanori Ojima , Miyu Takeuchi , Hiyori Takahashi , Makoto Ihara , David B. Sattelle , Kazuhiko Matsuda

Studies to date show that certain orthosteric binding sites are involved in the selective actions of neonicotinoids on insect nicotinic acetylcholine receptors (nAChRs). However, few investigations have utilized functional nAChRs consisting only of insect nAChR subunits. For the mechanism of selective neonicotinoid actions, loop C has been shown to play a role, whereas functional studies have advocated roles for loop D and loop G which appear to contribute more strongly to neonicotinoid actions than loop C. Here we have investigated the effects of P242E and P242S mutations in loop C of the Dα2 subunit and R81T mutation in loop D of the Dβ1 subunit on agonist actions of imidacloprid and thiacloprid for Drosophila melanogaster Dα1/Dα2/Dβ1 and Dα1/Dα2/Dβ1/Dβ2 nAChRs coexpressed with cofactors DmNACHO, DmRIC-3, DmTMX3 and DmUNC-50 in Xenopus laevis oocytes. The P242E and P242S mutations in loop C hardly affected the agonist affinity and efficacy of the neonicotinoids. In contrast, the R81T mutation in loop D decreased the affinity and efficacy of imidacloprid while substantially reducing the efficacy of thiacloprid. Combined loop C and loop D mutations resulted in further reduced efficacy of thiacloprid while having no such effect on imidacloprid actions. These results suggest that the proline in loop C and the arginine in loop D underly the binding of neonicotinoids with the greater contribution coming from loop D, and that thiacloprid relies less on the interactions with the arginine in loop D than imidacloprid, and hence is less susceptible to the R81T mutation in the development of resistance.

迄今为止的研究表明，某些正位结合位点参与了新烟碱类对昆虫烟碱乙酰胆碱受体（nAChRs）的选择性作用。然而，很少有研究利用仅由昆虫nAChR亚基组成的功能性nAChR。对于选择性新烟碱作用的机制，环C已被证明发挥了作用，在此，我们研究了Dα2亚基C环P242E和P242S突变和Dβ1亚基D环R81T突变对吡虫啉和噻虫啉对黑腹果蝇Dα1/Dα2/Dβ1和Dα1/Dα2/Dβ1/Dβ2 nAChRs与辅助因子DmNACHO、DmRIC-3、DmTMX3和DmUNC-50共表达的nAChRs在非洲爪蟾卵细胞中的受体活性的影响。环C的P242E和P242S突变几乎不影响新烟碱类激动剂的亲和力和药效。相反，环D中的R81T突变降低了吡虫啉的亲和力和药效，同时大幅降低了噻虫啉的药效。环C和环D的联合突变导致噻虫啉的药效进一步降低，而对吡虫啉的作用没有影响。这些结果表明，C环上的脯氨酸和D环上的精氨酸是新烟碱结合的基础，其中D环的贡献更大，与吡虫啉相比，噻虫啉对D环上精氨酸的相互作用依赖更小，因此在抗性发展过程中对R81T突变的敏感性更低。

{"title":"The functional impact of mutations in orthosteric binding site of the Drosophila melanogaster Dα2-containing nicotinic receptors points to a greater contribution to neonicotinoid selectivity of an arginine in loop D of Dβ1 than an introduced serine in loop C of Dα2","authors":"Ryo Ito , Hisanori Ojima , Miyu Takeuchi , Hiyori Takahashi , Makoto Ihara , David B. Sattelle , Kazuhiko Matsuda","doi":"10.1016/j.ibmb.2025.104380","DOIUrl":"10.1016/j.ibmb.2025.104380","url":null,"abstract":"<div><div>Studies to date show that certain orthosteric binding sites are involved in the selective actions of neonicotinoids on insect nicotinic acetylcholine receptors (nAChRs). However, few investigations have utilized functional nAChRs consisting only of insect nAChR subunits. For the mechanism of selective neonicotinoid actions, loop C has been shown to play a role, whereas functional studies have advocated roles for loop D and loop G which appear to contribute more strongly to neonicotinoid actions than loop C. Here we have investigated the effects of P242E and P242S mutations in loop C of the Dα2 subunit and R81T mutation in loop D of the Dβ1 subunit on agonist actions of imidacloprid and thiacloprid for <em>Drosophila melanogaster</em> Dα1/Dα2/Dβ1 and Dα1/Dα2/Dβ1/Dβ2 nAChRs coexpressed with cofactors DmNACHO, DmRIC-3, DmTMX3 and DmUNC-50 in <em>Xenopus laevis</em> oocytes. The P242E and P242S mutations in loop C hardly affected the agonist affinity and efficacy of the neonicotinoids. In contrast, the R81T mutation in loop D decreased the affinity and efficacy of imidacloprid while substantially reducing the efficacy of thiacloprid. Combined loop C and loop D mutations resulted in further reduced efficacy of thiacloprid while having no such effect on imidacloprid actions. These results suggest that the proline in loop C and the arginine in loop D underly the binding of neonicotinoids with the greater contribution coming from loop D, and that thiacloprid relies less on the interactions with the arginine in loop D than imidacloprid, and hence is less susceptible to the R81T mutation in the development of resistance.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"183 ","pages":"Article 104380"},"PeriodicalIF":3.7,"publicationDate":"2025-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144803118","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

Sterol-modified plants reduce aphid performance by limiting sterol availability 甾醇改性植物通过限制甾醇的可用性来降低蚜虫的表现。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-08-06 DOI: 10.1016/j.ibmb.2025.104382

Ivy W. Chen , Jiaxin Lei , Zekun Jin , Adriana E. Alvarez , Robert J. Grebenok , Stephen Fletcher , Spencer T. Behmer , Keyan Zhu-Salzman

Sterols are essential for eukaryotic cell structure and metabolism, yet insects cannot synthesize them de novo and must acquire them through their diet. For insect herbivores, plant-derived sterols are typically converted into cholesterol to support development and reproduction. We previously engineered Arabidopsis thaliana lines with silenced HYD1, resulting in altered sterol composition. In this study, we evaluated the performance of the phloem-feeding aphid Myzus persicae on these sterol-modified plants. Aphids reared on the modified lines (HYD1_RNAi10, 12, 25) exhibited significantly reduced growth, reproduction, and survival compared to those on wild-type Col-0. However, choice assays and electropenetrography (EPG) revealed no differences in host preference or probing and feeding behaviors. Unlike the chewing insect Plutella xylostella, M. persicae did not accumulate atypical sterols but instead showed a significant reduction of total sterol content. Phloem-sap analysis mirrored aphid sterol profiles, lacking the atypical sterols found in leaf tissue of the modified lines. RNA-seq of HYD1_RNAi lines revealed no induction of known plant defense pathways; instead, genes involved in translation and nitrate metabolism were upregulated. These findings show that HYD1 silencing reduces host suitability for aphids by limiting sterol availability for insect development and reproduction. Our results highlight the potential of sterol-modified plants as a promising strategy for managing phloem sap-feeding insect pests.

甾醇是真核细胞结构和代谢所必需的，但昆虫不能从头合成它们，必须通过饮食获得它们。对于食草昆虫来说，植物衍生的甾醇通常会转化为胆固醇来支持发育和繁殖。我们之前用沉默的HYD1基因改造拟南芥，导致甾醇成分发生改变。在本研究中，我们评估了韧皮部取食蚜虫对这些甾醇改性植物的作用。在改良系（HYD1RNAi10、12、25）上饲养的蚜虫的生长、繁殖和存活率明显低于野生型Col-0。然而，选择分析和电渗透（EPG）显示，寄主偏好或探测和摄食行为没有差异。与咀嚼昆虫小菜蛾不同，桃蚜不积累非典型甾醇，但总甾醇含量显著降低。韧皮部汁液分析反映了蚜虫的固醇谱，缺乏在改良系叶片组织中发现的非典型固醇。HYD1RNAi系的RNA-seq显示未诱导已知的植物防御途径；相反，参与翻译和硝酸盐代谢的基因被上调。这些发现表明，HYD1沉默通过限制昆虫发育和繁殖所需的甾醇来降低寄主对蚜虫的适应性。我们的研究结果强调了甾醇修饰植物作为一种有前途的策略来管理韧皮部取食汁液的害虫的潜力。

{"title":"Sterol-modified plants reduce aphid performance by limiting sterol availability","authors":"Ivy W. Chen , Jiaxin Lei , Zekun Jin , Adriana E. Alvarez , Robert J. Grebenok , Stephen Fletcher , Spencer T. Behmer , Keyan Zhu-Salzman","doi":"10.1016/j.ibmb.2025.104382","DOIUrl":"10.1016/j.ibmb.2025.104382","url":null,"abstract":"<div><div>Sterols are essential for eukaryotic cell structure and metabolism, yet insects cannot synthesize them <em>de novo</em> and must acquire them through their diet. For insect herbivores, plant-derived sterols are typically converted into cholesterol to support development and reproduction. We previously engineered <em>Arabidopsis thaliana</em> lines with silenced <em>HYD1</em>, resulting in altered sterol composition. In this study, we evaluated the performance of the phloem-feeding aphid <em>Myzus persicae</em> on these sterol-modified plants. Aphids reared on the modified lines (<em>HYD1</em><sub>RNAi</sub>10, 12, 25) exhibited significantly reduced growth, reproduction, and survival compared to those on wild-type Col-0. However, choice assays and electropenetrography (EPG) revealed no differences in host preference or probing and feeding behaviors. Unlike the chewing insect <em>Plutella xylostella</em>, <em>M. persica</em><em>e</em> did not accumulate atypical sterols but instead showed a significant reduction of total sterol content. Phloem-sap analysis mirrored aphid sterol profiles, lacking the atypical sterols found in leaf tissue of the modified lines. RNA-seq of <em>HYD1</em><sub>RNAi</sub> lines revealed no induction of known plant defense pathways; instead, genes involved in translation and nitrate metabolism were upregulated. These findings show that <em>HYD1</em> silencing reduces host suitability for aphids by limiting sterol availability for insect development and reproduction. Our results highlight the potential of sterol-modified plants as a promising strategy for managing phloem sap-feeding insect pests.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"183 ","pages":"Article 104382"},"PeriodicalIF":3.7,"publicationDate":"2025-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144803117","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

The transcriptomics of Hyphantria cunea salivary gland reveals its function in host plant adaptation 加利福尼亚棘球蚴唾液腺转录组学揭示了其在寄主植物适应中的功能

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-08-05 DOI: 10.1016/j.ibmb.2025.104379

Yajun Wang , Jiangyan Xie , Yansheng Sun , Shuming Cui , Jie Zhang , Qiaoya Zhang , Guocai Zhang , Bing Bi

Insects' salivary glands and their secretions play a pivotal role in their adaptation to host plants. Hyphantria cunea, a significant pest of both agriculture and forestry, feeds on a variety of host plants, yet the specific functions of its salivary glands in this adaptation process remain largely unclear. In this study, we compared the adaptability of fifth-instar larvae to Populus davidiana × P. bolleana (PDB) and artificial diet (AD). Through transcriptome analysis, 1439 differentially expressed genes (DEGs) were identified in the salivary glands of fifth-instar larvae feeding on PDB and AD. These DEGs include genes encoding various digestive and detoxification enzymes, which are enriched in pathways related to salivary secretion, digestion, and drug metabolism. Compared to larvae fed AD, the majority of digestive and detoxification enzyme genes were upregulated following consumption of PDB. Furthermore, the larvae enhanced the activities of two digestive enzymes (α-amylase and lipase) and four detoxification enzymes (cytochrome P450 monooxygenase, carboxylesterase, glutathione-S-transferase, and UDP-glycosyltransferases) in their salivary glands, thereby digesting the nutrients in the leaves while detoxifying the secondary metabolites contained within them. Silencing of CYP9E2 significantly reduced larval food intake and weight gain, and prolonged larval developmental duration. Taken together, our study identifies the salivary glands of H. cunea larvae as a critical tissue for executing digestive and detoxification functions, enhances understanding of the larval adaptability to host plants via salivary glands, and provides valuable insights for managing H. cunea infestations.

昆虫的唾液腺及其分泌物在它们适应寄主植物的过程中起着关键作用。美国棘球蚴（Hyphantria cunea）是农业和林业的一种重要害虫，它以多种寄主植物为食，但其唾液腺在这一适应过程中的具体功能仍不清楚。本研究比较了大叶杨（Populus davidiana × P. bolleana， PDB）和人工饲料（artificial diet， AD）对5龄幼虫的适应性。通过转录组分析，在取食PDB和AD的五龄幼虫的唾液腺中鉴定出1439个差异表达基因（DEGs）。这些deg包括编码各种消化和解毒酶的基因，这些基因在唾液分泌、消化和药物代谢的相关途径中富集。与饲喂AD的幼虫相比，食用PDB后，大部分消化酶和解毒酶基因上调。此外，幼虫提高了唾液腺中2种消化酶（α-淀粉酶和脂肪酶）和4种解毒酶（细胞色素P450单加氧酶、羧酯酶、谷胱甘肽- s -转移酶和udp -糖基转移酶）的活性，从而消化叶片中的营养物质，同时对叶片中的次生代谢物进行解毒。CYP9E2的沉默显著降低了幼虫的食物摄取量和体重增加，延长了幼虫的发育时间。综上所述，我们的研究确定了美洲锥虫幼虫的唾液腺是执行消化和解毒功能的关键组织，增强了对幼虫通过唾液腺对寄主植物适应性的认识，并为管理美洲锥虫侵染提供了有价值的见解。

{"title":"The transcriptomics of Hyphantria cunea salivary gland reveals its function in host plant adaptation","authors":"Yajun Wang , Jiangyan Xie , Yansheng Sun , Shuming Cui , Jie Zhang , Qiaoya Zhang , Guocai Zhang , Bing Bi","doi":"10.1016/j.ibmb.2025.104379","DOIUrl":"10.1016/j.ibmb.2025.104379","url":null,"abstract":"<div><div>Insects' salivary glands and their secretions play a pivotal role in their adaptation to host plants. <em>Hyphantria cunea</em>, a significant pest of both agriculture and forestry, feeds on a variety of host plants, yet the specific functions of its salivary glands in this adaptation process remain largely unclear. In this study, we compared the adaptability of fifth-instar larvae to <em>Populus davidiana</em> × <em>P. bolleana</em> (PDB) and artificial diet (AD). Through transcriptome analysis, 1439 differentially expressed genes (DEGs) were identified in the salivary glands of fifth-instar larvae feeding on PDB and AD. These DEGs include genes encoding various digestive and detoxification enzymes, which are enriched in pathways related to salivary secretion, digestion, and drug metabolism. Compared to larvae fed AD, the majority of digestive and detoxification enzyme genes were upregulated following consumption of PDB. Furthermore, the larvae enhanced the activities of two digestive enzymes (α-amylase and lipase) and four detoxification enzymes (cytochrome P450 monooxygenase, carboxylesterase, glutathione-S-transferase, and UDP-glycosyltransferases) in their salivary glands, thereby digesting the nutrients in the leaves while detoxifying the secondary metabolites contained within them. Silencing of <em>CYP9E2</em> significantly reduced larval food intake and weight gain, and prolonged larval developmental duration. Taken together, our study identifies the salivary glands of <em>H</em>. <em>cunea</em> larvae as a critical tissue for executing digestive and detoxification functions, enhances understanding of the larval adaptability to host plants via salivary glands, and provides valuable insights for managing <em>H</em>. <em>cunea</em> infestations.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"183 ","pages":"Article 104379"},"PeriodicalIF":3.7,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144771164","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

20-Hydroxyecdysone regulates the expression of antimicrobial peptides through dorsal and relish in Helicoverpa armigera 20-羟基蜕皮激素通过棉铃虫背侧和津津乐道调节抗菌肽的表达。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-07-30 DOI: 10.1016/j.ibmb.2025.104378

Caihua Chen , Youying Yan , Jiaxin Wang , Lizhen Zhou , Zhiqiang Lu

20-hydroxyecdysone (20E) plays a key role in insect development, not only regulating molting and metamorphosis but also participating in the innate immune responses. The regulation of immunity by 20E varies among different insect species. The pathway and mechanism of 20E regulates immunity in the cotton bollworm (Helicoverpa armigera) remain unclear. In this study, we confirmed that the enhanced immunity is regulated by 20E in H. armigera. 20E upregulates the expression of antimicrobial peptides (AMPs). Knockdown of Dorsal suppressed the 20E-induced expression of AMPs, while knockdown of Relish similarly inhibited the expression of AMPs except Moricin. Overexpression of Dorsal promotes its nuclear translocation and activates the expression of AMPs. However, overexpression of Cactus (IκB-like protein, the inhibitor of NFκB) isoform A binds to Dorsal and inhibits its nuclear translocation. 20E upregulates Dorsal expression via direct EcR mediation and facilitates Dorsal release through phosphorylation of Cactus A. We also report spontaneous activation of HaRelish through auto-proteolytic cleavage, liberating its N-terminal domain (Rel-N). Nuclear Rel-N interacts with Dorsal to co-regulate AMP transcription. These data indicate that 20E induces the expression of AMPs in H. armigera through activation of Dorsal and Relish, thereby enhancing immunity during the metamorphosis stage.

20-羟基蜕皮激素（20-hydroxyecdysone, 20E）在昆虫发育过程中起着关键作用，不仅调节昆虫的蜕皮和变态，还参与昆虫的先天免疫反应。20E对不同昆虫的免疫调节是不同的。20E调节棉铃虫（Helicoverpa armigera）免疫的途径和机制尚不清楚。在本研究中，我们证实了这种增强的免疫是由20E调节的。20E上调抗菌肽（AMPs）的表达。低敲Dorsal抑制了20e诱导的AMPs的表达，而低敲enjoy同样抑制了除Moricin外的AMPs的表达。Dorsal的过表达促进其核易位，激活amp的表达。然而，过表达Cactus （i - κ b样蛋白，nf - κ b抑制剂）异构体A与背侧结合并抑制其核易位。20E通过直接EcR介导上调背侧表达，并通过Cactus a的磷酸化促进背侧释放。我们还报道了ha回味蛋白通过自身蛋白水解裂解而自发激活，释放其n端结构域（Rel-N）。核Rel-N与Dorsal相互作用，共同调节AMP转录。这些数据表明，20E通过激活Dorsal和回味蛋白诱导棉铃虫AMPs的表达，从而增强棉铃虫变态期的免疫力。

{"title":"20-Hydroxyecdysone regulates the expression of antimicrobial peptides through dorsal and relish in Helicoverpa armigera","authors":"Caihua Chen , Youying Yan , Jiaxin Wang , Lizhen Zhou , Zhiqiang Lu","doi":"10.1016/j.ibmb.2025.104378","DOIUrl":"10.1016/j.ibmb.2025.104378","url":null,"abstract":"<div><div>20-hydroxyecdysone (20E) plays a key role in insect development, not only regulating molting and metamorphosis but also participating in the innate immune responses. The regulation of immunity by 20E varies among different insect species. The pathway and mechanism of 20E regulates immunity in the cotton bollworm (<em>Helicoverpa armigera</em>) remain unclear. In this study, we confirmed that the enhanced immunity is regulated by 20E in <em>H. armigera</em>. 20E upregulates the expression of antimicrobial peptides (AMPs). Knockdown of Dorsal suppressed the 20E-induced expression of AMPs, while knockdown of Relish similarly inhibited the expression of AMPs except <em>Moricin</em>. Overexpression of Dorsal promotes its nuclear translocation and activates the expression of AMPs. However, overexpression of Cactus (IκB-like protein, the inhibitor of NFκB) isoform A binds to Dorsal and inhibits its nuclear translocation. 20E upregulates Dorsal expression via direct EcR mediation and facilitates Dorsal release through phosphorylation of Cactus A. We also report spontaneous activation of HaRelish through auto-proteolytic cleavage, liberating its N-terminal domain (Rel-N). Nuclear Rel-N interacts with Dorsal to co-regulate AMP transcription. These data indicate that 20E induces the expression of AMPs in <em>H. armigera</em> through activation of Dorsal and Relish, thereby enhancing immunity during the metamorphosis stage.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"183 ","pages":"Article 104378"},"PeriodicalIF":3.7,"publicationDate":"2025-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144764262","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

The arthropod P450 Enchiridion: An integrated web resource for research on P450s 节肢动物P450百科：一个用于研究P450的综合网络资源。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-07-24 DOI: 10.1016/j.ibmb.2025.104377

Jason Charamis , Wannes Dermauw , Thomas Van Leeuwen , John Vontas , René Feyereisen

We present the Arthropod P450 Enchiridion (arthropodp450.eu), a website based on the DokuWiki architecture and designed to provide resources on structures and functions of CYP genes and P450 enzymes from the most diverse and numerous animals, the arthropods. Interlinked pages cover some essential common features of P450 enzymes from rules of nomenclature to enzyme mechanism and protein structure. Specific to arthropods, lookup tables allow the rapid assignment of currently 1308 CYP families to their nine respective CYP clans, as well as the distribution of P450s from full CYPomes into clans. Manually annotated CYPomes from currently 75 species are presented with their correct CYP names as well as nearly 3000 full-length, named insect transcripts. The Arthropod P450 Enchiridion offers a BLAST function addressing over 11,000 accurately named P450 protein sequences. P450 functions as assessed by in vitro heterologous expression, RNAi phenotype, transgenic in vivo expression, or the phenotype of null mutants are presented for over 1000 cases. The data show as expected a focus on insect and mite pest species, but also highlights the remaining gaps in our knowledge of sequence and function of P450s in less studied groups of arthropods. The architecture of the DokuWiki enables regular updates and expansions to which authors are welcome to contribute.

我们介绍了节肢动物P450 Enchiridion (arthropodp450.eu)，这是一个基于DokuWiki架构的网站，旨在提供来自最多样化和数量最多的动物节肢动物的CYP基因和P450酶的结构和功能资源。相互链接的页面涵盖了P450酶的一些基本共同特征，从命名规则到酶的机制和蛋白质结构。具体到节肢动物，查找表允许将目前1308个CYP家族快速分配到9个各自的CYP氏族，以及从完整cypome到氏族的p450分布。人工标注了目前75个物种的cypome，提供了正确的CYP名称以及近3000个全长的命名昆虫转录本。节肢动物P450 Enchiridion提供BLAST功能，定位超过11,000个精确命名的P450蛋白序列。通过体外异源表达、RNAi表型、转基因体内表达或无突变体表型来评估P450功能的案例超过1000例。这些数据如预期的那样显示了对昆虫和螨虫害虫物种的关注，但也突出了我们对较少研究的节肢动物群体中p450的序列和功能的了解仍然存在空白。DokuWiki的体系结构支持定期更新和扩展，欢迎作者对此做出贡献。

{"title":"The arthropod P450 Enchiridion: An integrated web resource for research on P450s","authors":"Jason Charamis , Wannes Dermauw , Thomas Van Leeuwen , John Vontas , René Feyereisen","doi":"10.1016/j.ibmb.2025.104377","DOIUrl":"10.1016/j.ibmb.2025.104377","url":null,"abstract":"<div><div>We present the Arthropod P450 Enchiridion (<span><span>arthropodp450.eu</span><svg><path></path></svg></span>), a website based on the DokuWiki architecture and designed to provide resources on structures and functions of CYP genes and P450 enzymes from the most diverse and numerous animals, the arthropods. Interlinked pages cover some essential common features of P450 enzymes from rules of nomenclature to enzyme mechanism and protein structure. Specific to arthropods, lookup tables allow the rapid assignment of currently 1308 CYP families to their nine respective CYP clans, as well as the distribution of P450s from full CYPomes into clans. Manually annotated CYPomes from currently 75 species are presented with their correct CYP names as well as nearly 3000 full-length, named insect transcripts. The Arthropod P450 Enchiridion offers a BLAST function addressing over 11,000 accurately named P450 protein sequences. P450 functions as assessed by <em>in vitro</em> heterologous expression, RNAi phenotype, transgenic <em>in vivo</em> expression, or the phenotype of null mutants are presented for over 1000 cases. The data show as expected a focus on insect and mite pest species, but also highlights the remaining gaps in our knowledge of sequence and function of P450s in less studied groups of arthropods. The architecture of the DokuWiki enables regular updates and expansions to which authors are welcome to contribute.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"183 ","pages":"Article 104377"},"PeriodicalIF":3.7,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144717181","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

Functional analysis of vesicular glutamate transporter in Chilo suppressalis: Implications for glutamate transport and pest management 抑制辣椒水疱谷氨酸转运蛋白的功能分析：谷氨酸转运和害虫管理的意义。

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

Insect Biochemistry and Molecular Biology

Pub Date : 2025-07-23 DOI: 10.1016/j.ibmb.2025.104376

Xin Mao , Yichi Zhang , Qiwen Pu , Ali Hasnain , Xinyu Liu , Guanghua Luo , Chunqing Zhao

Glutamate is the primary excitatory neurotransmitter in animals, and its storage and release are regulated by vesicular glutamate transporters (vGluTs) in mammals. However, the function of vGluTs in agricultural pests remains largely unexplored. In this study, we successfully cloned the full-length vGluT gene from the rice striped stem borer (RSB) Chilo suppressalis, a major rice pest. Multiple sequence alignment revealed that the amino acid sequence of CsvGluT shared more than 69.82 % similarity with other known insect vGluTs. In the phylogenetic tree, CsvGluT was clustered into a clade with the Asian corn borer Ostrinia furnacalis, which also belongs to Crambidae family. Spatial-temporal expression analysis showed that CsvGluT was highly expressed in the larval heads and abundant in the 1^st- and 2^nd-instar larvae, whose expression levels are more than three-fold higher than those in the 3^rd- to 6^th -instar larvae. Electrophysiological recordings indicated that CsvGluT exhibited functional properties like mammalian vGluTs under high concentrations of protons and chloride ions. Furthermore, inhibitors of mammalian vGluTs, including evans blue (EB) and 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid (DIDS), significantly inhibited CsvGluT activity. RNA interference (RNAi)-mediated knockdown of CsvGluT significantly reduced the survival rate of RSB to 66 %, highlighting its physiological role and a potential target for RNAi-based RSB control. In summary, our findings elucidated the physiological function of vGluT in RSB and provide a theoretical foundation for advanced research in the development of RNAi-based transporter-targeted strategies for pest management.

谷氨酸是动物主要的兴奋性神经递质，其储存和释放受哺乳动物水疱性谷氨酸转运蛋白（vGluTs）的调控。然而，vglut在农业害虫中的作用在很大程度上仍未被探索。在本研究中，我们成功克隆了水稻螟虫（RSB） Chilo suppressalis的全长vGluT基因。多序列比对结果表明，CsvGluT与其他已知昆虫vglut的氨基酸序列相似性超过69.82%。在系统发育树上，CsvGluT与亚洲玉米螟（Ostrinia furnacalis）聚为一个分支，后者也属于玉米螟科。时空表达分析表明，CsvGluT在幼虫头部高度表达，在1龄和2龄幼虫中表达量丰富，表达量是3 ~ 6龄幼虫的3倍以上。电生理记录表明，CsvGluT在高浓度质子和氯离子作用下表现出与哺乳动物vglut类似的功能特性。此外，哺乳动物vglut的抑制剂，包括evans蓝（EB）和4,4'-二异硫氰基二苯乙烯-2,2'-二磺酸（DIDS），显著抑制了CsvGluT的活性。RNA干扰（RNAi）介导的CsvGluT敲低可显著降低RSB的存活率至66%，突出了其生理作用和基于RNAi的RSB控制的潜在靶点。综上所述，我们的研究结果阐明了vGluT在RSB中的生理功能，为进一步研究基于rnai的转运蛋白靶向害虫防治策略提供了理论基础。

{"title":"Functional analysis of vesicular glutamate transporter in Chilo suppressalis: Implications for glutamate transport and pest management","authors":"Xin Mao , Yichi Zhang , Qiwen Pu , Ali Hasnain , Xinyu Liu , Guanghua Luo , Chunqing Zhao","doi":"10.1016/j.ibmb.2025.104376","DOIUrl":"10.1016/j.ibmb.2025.104376","url":null,"abstract":"<div><div>Glutamate is the primary excitatory neurotransmitter in animals, and its storage and release are regulated by vesicular glutamate transporters (vGluTs) in mammals. However, the function of vGluTs in agricultural pests remains largely unexplored. In this study, we successfully cloned the full-length <em>vGluT</em> gene from the rice striped stem borer (RSB) <em>Chilo suppressalis</em>, a major rice pest. Multiple sequence alignment revealed that the amino acid sequence of <em>Cs</em>vGluT shared more than 69.82 % similarity with other known insect vGluTs. In the phylogenetic tree, <em>Cs</em>vGluT was clustered into a clade with the Asian corn borer <em>Ostrinia furnacalis</em>, which also belongs to Crambidae family. Spatial-temporal expression analysis showed that <em>CsvGluT</em> was highly expressed in the larval heads and abundant in the 1<sup>st</sup>- and 2<sup>nd</sup>-instar larvae, whose expression levels are more than three-fold higher than those in the 3<sup>rd</sup>- to 6<sup>th</sup> -instar larvae. Electrophysiological recordings indicated that <em>Cs</em>vGluT exhibited functional properties like mammalian vGluTs under high concentrations of protons and chloride ions. Furthermore, inhibitors of mammalian vGluTs, including evans blue (EB) and 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid (DIDS), significantly inhibited <em>Cs</em>vGluT activity. RNA interference (RNAi)-mediated knockdown of <em>CsvGluT</em> significantly reduced the survival rate of RSB to 66 %, highlighting its physiological role and a potential target for RNAi-based RSB control. In summary, our findings elucidated the physiological function of vGluT in RSB and provide a theoretical foundation for advanced research in the development of RNAi-based transporter-targeted strategies for pest management.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"183 ","pages":"Article 104376"},"PeriodicalIF":3.7,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144717180","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