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

Structural characterization and proteomic profiling of oviposition secretions across three rice planthopper species 三种稻飞虱产卵分泌物的结构特征和蛋白质组分析

IF 3.2 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2024-11-22 DOI: 10.1016/j.ibmb.2024.104220

Jia-Bao Lu , Peng-Peng Ren , Ying Tian , Yan-Yan Yang , Qing-Kai Feng , Xiao-Ya Zhang , Fang He , Hai-Jian Huang , Jian-Ping Chen , Jun-Min Li , Chuan-Xi Zhang

Insect oviposition secretions play crucial roles during the reproductive process, yet systematic studies on their structural characterization and protein compositions remain limited. This study investigated the oviposition secretions of three major rice pests: the brown planthopper (Nilaparvata lugens, BPH), small brown planthopper (Laodelphax striatella, SBPH), and white-backed planthopper (Sogatella furcifera, WBPH). Ultrastructural observation revealed differences in the oviposition secretions of them. The eggs of BPH and SBPH were adhered to rice tissue by abundant secretions, while WBPH eggs were embedded deeper within the leaf sheath with less secretions. Proteomic analysis identified 111, 98, and 66 oviposition secretion proteins (OSPs) in BPH, SBPH, and WBPH, respectively. 4 common protein subgroups were shared among them, along with varying numbers of shared subgroups between species pairs. Notably, the majority of OSPs were exclusively found in one species, indicating the existence of both similar and specialized functions unique to each planthopper species. The functions of 4 uncharacterized OSPs (Nl.chr07.0363, Nl.chr12.078, Nl.chr11.716, Nl.scaffold.0714) that were uniquely identified in the BPH were studied by maternal RNAi. Downregulation of each of these 4 protein-coding genes led to a significant decrease in egg production and hatchability. Moreover, knockdown of Nl.chr12.078 or Nl.chr07.0363 also disrupt the secretory function of the lateral oviduct. In conclusion, this study provides insights into the structural characteristics and protein components of the oviposition secretions of BPH, SBPH, and WBPH, which could serve as potential targets for RNAi-based pest control and lay a foundation for future studies on insect-plant interactions mediated by oviposition secretions.

昆虫的产卵分泌物在繁殖过程中起着至关重要的作用，但对其结构特征和蛋白质组成的系统研究仍然有限。本研究调查了三种主要水稻害虫的产卵分泌物：褐跳甲（Nilaparvata lugens，BPH）、小褐跳甲（Laodelphax striatella，SBPH）和白背跳甲（Sogatella furcifera，WBPH）。超微结构观察发现了它们产卵分泌物的差异。BPH和SBPH的卵被大量分泌物粘附在水稻组织上，而WBPH的卵则深埋在叶鞘中，分泌物较少。蛋白质组分析在 BPH、SBPH 和 WBPH 中分别发现了 111、98 和 66 个产卵分泌蛋白（OSPs）。它们之间共有 4 个共同的蛋白质亚群，物种对之间也有不同数量的共同亚群。值得注意的是，大多数 OSPs 只存在于一个物种中，这表明每个跳虫物种既有相似的功能，也有其特有的功能。研究人员通过母体 RNAi 研究了 4 个未表征的 OSPs（Nl.chr07.0363、Nl.chr12.078、Nl.chr11.716 和 Nl.scaffold.0714）的功能，这 4 个 OSPs 是在 BPH 中唯一发现的。下调这4个蛋白编码基因会导致产卵量和孵化率显著下降。此外，敲除Nl.chr12.078或Nl.chr07.0363也会破坏侧输卵管的分泌功能。总之，本研究深入揭示了BPH、SBPH和WBPH产卵分泌物的结构特征和蛋白质成分，可作为基于RNAi的害虫控制的潜在靶标，并为今后研究由产卵分泌物介导的昆虫与植物之间的相互作用奠定了基础。

{"title":"Structural characterization and proteomic profiling of oviposition secretions across three rice planthopper species","authors":"Jia-Bao Lu , Peng-Peng Ren , Ying Tian , Yan-Yan Yang , Qing-Kai Feng , Xiao-Ya Zhang , Fang He , Hai-Jian Huang , Jian-Ping Chen , Jun-Min Li , Chuan-Xi Zhang","doi":"10.1016/j.ibmb.2024.104220","DOIUrl":"10.1016/j.ibmb.2024.104220","url":null,"abstract":"<div><div>Insect oviposition secretions play crucial roles during the reproductive process, yet systematic studies on their structural characterization and protein compositions remain limited. This study investigated the oviposition secretions of three major rice pests: the brown planthopper (<em>Nilaparvata lugens</em>, BPH), small brown planthopper (<em>Laodelphax striatella</em>, SBPH), and white-backed planthopper (<em>Sogatella furcifera</em>, WBPH). Ultrastructural observation revealed differences in the oviposition secretions of them. The eggs of BPH and SBPH were adhered to rice tissue by abundant secretions, while WBPH eggs were embedded deeper within the leaf sheath with less secretions. Proteomic analysis identified 111, 98, and 66 oviposition secretion proteins (OSPs) in BPH, SBPH, and WBPH, respectively. 4 common protein subgroups were shared among them, along with varying numbers of shared subgroups between species pairs. Notably, the majority of OSPs were exclusively found in one species, indicating the existence of both similar and specialized functions unique to each planthopper species. The functions of 4 uncharacterized OSPs (Nl.chr07.0363, Nl.chr12.078, Nl.chr11.716, Nl.scaffold.0714) that were uniquely identified in the BPH were studied by maternal RNAi. Downregulation of each of these 4 protein-coding genes led to a significant decrease in egg production and hatchability. Moreover, knockdown of <em>Nl.chr12.078</em> or <em>Nl.chr07.0363</em> also disrupt the secretory function of the lateral oviduct. In conclusion, this study provides insights into the structural characteristics and protein components of the oviposition secretions of BPH, SBPH, and WBPH, which could serve as potential targets for RNAi-based pest control and lay a foundation for future studies on insect-plant interactions mediated by oviposition secretions.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"176 ","pages":"Article 104220"},"PeriodicalIF":3.2,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142699016","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 immune function of thioester-containing proteins in typical invertebrate disease vectors 典型无脊椎动物病媒中含硫酯的蛋白质的免疫功能。

IF 3.2 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2024-11-22 DOI: 10.1016/j.ibmb.2024.104218

Qianqian Zhang , Xia Zhou , Tingting Feng , Hao Tong , Jun Wang , Jianfeng Dai

Disease vectors, such as arthropods, primarily rely on innate immunity to counteract pathogen invasions, typically through the recognition and binding of pathogen-associated molecular patterns (PAMPs) by the host's pattern recognition receptors (PRRs). As a conserved immune effector gene family from insects to mammals, the complement system may play an essential role in combating pathogenic microorganisms. In arthropods, the complement proteins are often referred to as thioester-containing proteins (TEPs) because thioester motifs are one of the essential functional domains of the first proteins characterized within the C3 and A₂M family. TEPs mainly function as specialized PRRs in sensing and binding to pathogens or their components. This paper presents a comprehensive review of the common domain and functions of TEPs in major disease vectors, in particular the specific decision-making ones expressed by Arthropoda (medical arthropods) and Mollusca (Biomphalaria glabrata) after pathogen infections. The relationship between the structure and antibacterial/antiviral activities of TEPs would further our understandings on the mechanisms governing the initiation of innate immune responses in typical disease vectors.

节肢动物等疾病载体主要依靠先天性免疫来抵御病原体的入侵，通常是通过宿主的模式识别受体（PRRs）识别和结合病原体相关分子模式（PAMPs）。作为一个从昆虫到哺乳动物的保守免疫效应基因家族，补体系统可能在抗击病原微生物的过程中发挥着至关重要的作用。在节肢动物中，补体蛋白通常被称为含硫代酯蛋白（TEPs），因为硫代酯基团是 C3 和 A2M 家族中首批表征蛋白的重要功能域之一。TEPs 主要在感知和结合病原体或其成分方面发挥专门的 PRRs 功能。本文全面综述了 TEPs 在主要疾病媒介中的共同结构域和功能，特别是节肢动物（医学节肢动物）和软体动物（玻璃鲎）在病原体感染后表达的特定决策性 TEPs。TEPs 的结构与抗菌/抗病毒活性之间的关系将进一步加深我们对典型病媒先天免疫反应启动机制的理解。

{"title":"The immune function of thioester-containing proteins in typical invertebrate disease vectors","authors":"Qianqian Zhang , Xia Zhou , Tingting Feng , Hao Tong , Jun Wang , Jianfeng Dai","doi":"10.1016/j.ibmb.2024.104218","DOIUrl":"10.1016/j.ibmb.2024.104218","url":null,"abstract":"<div><div>Disease vectors, such as arthropods, primarily rely on innate immunity to counteract pathogen invasions, typically through the recognition and binding of pathogen-associated molecular patterns (PAMPs) by the host's pattern recognition receptors (PRRs). As a conserved immune effector gene family from insects to mammals, the complement system may play an essential role in combating pathogenic microorganisms. In arthropods, the complement proteins are often referred to as thioester-containing proteins (TEPs) because thioester motifs are one of the essential functional domains of the first proteins characterized within the C3 and A<sub>2</sub>M family. TEPs mainly function as specialized PRRs in sensing and binding to pathogens or their components. This paper presents a comprehensive review of the common domain and functions of TEPs in major disease vectors, in particular the specific decision-making ones expressed by <em>Arthropoda</em> (medical arthropods) and <em>Mollusca</em> (<em>Biomphalaria glabrata</em>) after pathogen infections. The relationship between the structure and antibacterial/antiviral activities of TEPs would further our understandings on the mechanisms governing the initiation of innate immune responses in typical disease vectors.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"176 ","pages":"Article 104218"},"PeriodicalIF":3.2,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142694939","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

Suppression of the H3K27me3 demethylase disrupts diapause formation in mosquito Culex pipiens 抑制 H3K27me3 去甲基化酶会破坏库蚊的休眠形成。

IF 3.2 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2024-11-20 DOI: 10.1016/j.ibmb.2024.104216

Xueyan Wei , Prabin Dhungana , Kaylah Callender , Berhanu Zewde , Fu Chen , Sung Joon Kim , Cheolho Sim

Diapause (D) is a hormonally controlled alternative developmental pathway that allows mosquitoes to survive harsh winter conditions. Key characteristics of mosquito diapause include elevated lipid storage, enhanced stress and cold endurance, and extended longevity. These phenotypic changes are often associated with dynamic alterations in the transcriptome and epigenome. In our previous study, we identified significantly lower H3K27me2 levels in the fat body (FB) of diapausing Culex pipiens. However, the specific roles of the repressive H3K27 methylation marks in mosquito diapause have not been investigated. In the present study, we employed the effective histone lysine demethylase inhibitor GSK-J4 to assess the functions of H3K27me3 levels in the fat body on diapause initiation and phenotypes in Cx. pipiens. Results from solid-state NMR (ssNMR), Fourier-transform infrared spectroscopy (FTIR), and biochemical assays suggest that elevated H3K27me3 levels via GSK-J4 inhibition led to disrupted accumulation of lipids and glycogen in diapausing mosquitoes. GSK-J4 treatment also increased the mortality rate, resulting in lower survivability in treated mosquitoes. Together, these findings propose a crucial role for H3K27me3 in diapause formation, particularly related to energy metabolism. Our results provide a potential target for novel vector control strategies for this species.

休眠（D）是一种受激素控制的替代发育途径，可使蚊子在严冬条件下存活下来。蚊子休眠期的主要特征包括脂质储存增加、应激和耐寒能力增强以及寿命延长。这些表型变化往往与转录组和表观基因组的动态变化有关。在我们之前的研究中，我们发现会绝食的库蚊脂肪体（FB）中的 H3K27me2 水平明显较低。然而，抑制性 H3K27 甲基化标记在蚊子休眠中的具体作用尚未得到研究。在本研究中，我们采用了有效的组蛋白赖氨酸去甲基化酶抑制剂 GSK-J4 来评估脂肪体中 H3K27me3 水平对喙蚊休眠启动和表型的作用。固态核磁共振（ssNMR）、傅立叶变换红外光谱（FTIR）和生化测定的结果表明，通过抑制GSK-J4提高H3K27me3的水平，会导致暂停期蚊子体内脂质和糖原的堆积紊乱。GSK-J4 处理也增加了死亡率，导致处理过的蚊子存活率降低。总之，这些研究结果表明，H3K27me3 在昏睡形成过程中起着关键作用，特别是与能量代谢有关。我们的研究结果为该物种的新型病媒控制策略提供了一个潜在目标。

{"title":"Suppression of the H3K27me3 demethylase disrupts diapause formation in mosquito Culex pipiens","authors":"Xueyan Wei , Prabin Dhungana , Kaylah Callender , Berhanu Zewde , Fu Chen , Sung Joon Kim , Cheolho Sim","doi":"10.1016/j.ibmb.2024.104216","DOIUrl":"10.1016/j.ibmb.2024.104216","url":null,"abstract":"<div><div>Diapause (D) is a hormonally controlled alternative developmental pathway that allows mosquitoes to survive harsh winter conditions. Key characteristics of mosquito diapause include elevated lipid storage, enhanced stress and cold endurance, and extended longevity. These phenotypic changes are often associated with dynamic alterations in the transcriptome and epigenome. In our previous study, we identified significantly lower H3K27me2 levels in the fat body (FB) of diapausing <em>Culex pipiens</em>. However, the specific roles of the repressive H3K27 methylation marks in mosquito diapause have not been investigated. In the present study, we employed the effective histone lysine demethylase inhibitor GSK-J4 to assess the functions of H3K27me3 levels in the fat body on diapause initiation and phenotypes in <em>Cx. pipiens</em>. Results from solid-state NMR (ssNMR), Fourier-transform infrared spectroscopy (FTIR), and biochemical assays suggest that elevated H3K27me3 levels via GSK-J4 inhibition led to disrupted accumulation of lipids and glycogen in diapausing mosquitoes. GSK-J4 treatment also increased the mortality rate, resulting in lower survivability in treated mosquitoes. Together, these findings propose a crucial role for H3K27me3 in diapause formation, particularly related to energy metabolism. Our results provide a potential target for novel vector control strategies for this species.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"176 ","pages":"Article 104216"},"PeriodicalIF":3.2,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142692272","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 near-complete genome reveals the population evolution of the cotton-melon aphid Aphis gossypii 近乎完整的基因组揭示了棉瓜蚜 Aphis gossypii 的种群进化。

IF 3.2 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2024-11-19 DOI: 10.1016/j.ibmb.2024.104215

Tianxing Jing , Jin Yang , Jilong Pan , Xiaoning Liu , Xinyi Yang , Muhammad Farhan , Honghua Su , Xiaoyan Ma , Shuai Zhang

The cotton-melon aphid Aphis gossypii Glover is a severe pest worldwide. Interhaplotype genomic variation can be used as a starting point to analyze the adaptability of Ap. gossypii. In this study, we utilized long-read PacBio HiFi sequencing and HiC scaffolding techniques to assemble a near telomere-to-telomere gap-free genome assembly of Hap4. The assembly had two gaps totaling 321.24 Mb. We characterized five telomeric repetitive regions (GGTTA)_n, including the four found at the 3′ end of the chromosomes, and obtained new structural information about the telomeres. Due to the improved sequencing technology, we also identified more than 55.03 Mb of repetitive DNA in the genome assembly of Hap4, which contributed significantly to the increase in genome size compared to that of Hap1 and Hap3. Most of the additional repetitive DNA content was located on the X chromosome, and the tandem repeat sequence occupied 16.8% of the X chromosome length. The Hap4 assembly showed that the X chromosome exhibited a greater abundance of AT-rich satDNA arrays (11 satDNA arrays longer than 100 kb) than that observed in the autosomes (A1 and A2 harboured 3 and 1 satDNA arrays). We detected presence-absence variations, insertions, and deletions events between Hap1, Hap3, and Hap4 Ap. gossypii, which had significant effects on gene expression. Additionally, we identified a male-specific glyceraldehyde-3-phosphate dehydrogenase of fungal origin in all strains of Ap. gossypii. This comprehensive genome assembly provides valuable insights into the structural characteristics of highly repetitive regions and allows comparative genomic analyses that facilitate our understanding of Ap. gossypii's adaptation and diversification.

棉瓜蚜 Aphis gossypii Glover 是世界上一种严重的害虫。组间基因组变异可作为分析棉铃蚜适应性的起点。在这项研究中，我们利用长线程 PacBio HiFi 测序和 HiC 搭架技术，对 Hap4 进行了近端粒到端粒的无间隙基因组组装。该装配有两个缺口，总计 321.24 Mb。我们鉴定了五个端粒重复区（GGTTA）n，包括染色体 3' 端的四个重复区，并获得了端粒的新结构信息。由于测序技术的改进，我们还在 Hap4 的基因组组装中发现了超过 55.03 Mb 的重复性 DNA，与 Hap1 和 Hap3 相比，这大大增加了基因组的大小。大部分额外的重复 DNA 位于 X 染色体上，串联重复序列占据了 X 染色体长度的 16.8%。Hap4 组合显示，X 染色体上富含 AT 的 satDNA 阵列（11 个长度超过 100 kb 的 satDNA 阵列）比常染色体上的更多（A1 和 A2 分别含有 3 个和 1 个 satDNA 阵列）。我们检测到了 Hap1、Hap3 和 Hap4 Ap. gossypii 之间的存在-不存在变异、插入和缺失事件，它们对基因表达有显著影响。此外，我们还在所有 Ap. gossypii 菌株中发现了一种雄性特异性的 3-磷酸甘油醛脱氢酶。这一全面的基因组组装为我们深入了解高度重复区域的结构特征提供了宝贵的信息，并允许进行基因组比较分析，从而促进我们对棉铃虫适应性和多样性的了解。

{"title":"A near-complete genome reveals the population evolution of the cotton-melon aphid Aphis gossypii","authors":"Tianxing Jing , Jin Yang , Jilong Pan , Xiaoning Liu , Xinyi Yang , Muhammad Farhan , Honghua Su , Xiaoyan Ma , Shuai Zhang","doi":"10.1016/j.ibmb.2024.104215","DOIUrl":"10.1016/j.ibmb.2024.104215","url":null,"abstract":"<div><div>The cotton-melon aphid <em>Aphis gossypii</em> Glover is a severe pest worldwide. Interhaplotype genomic variation can be used as a starting point to analyze the adaptability of <em>Ap. gossypii</em>. In this study, we utilized long-read PacBio HiFi sequencing and HiC scaffolding techniques to assemble a near telomere-to-telomere gap-free genome assembly of Hap4. The assembly had two gaps totaling 321.24 Mb. We characterized five telomeric repetitive regions (GGTTA)<sub>n</sub>, including the four found at the 3′ end of the chromosomes, and obtained new structural information about the telomeres. Due to the improved sequencing technology, we also identified more than 55.03 Mb of repetitive DNA in the genome assembly of Hap4, which contributed significantly to the increase in genome size compared to that of Hap1 and Hap3. Most of the additional repetitive DNA content was located on the X chromosome, and the tandem repeat sequence occupied 16.8% of the X chromosome length. The Hap4 assembly showed that the X chromosome exhibited a greater abundance of AT-rich satDNA arrays (11 satDNA arrays longer than 100 kb) than that observed in the autosomes (A1 and A2 harboured 3 and 1 satDNA arrays). We detected presence-absence variations, insertions, and deletions events between Hap1, Hap3, and Hap4 <em>Ap. gossypii</em>, which had significant effects on gene expression. Additionally, we identified a male-specific glyceraldehyde-3-phosphate dehydrogenase of fungal origin in all strains of <em>Ap. gossypii</em>. This comprehensive genome assembly provides valuable insights into the structural characteristics of highly repetitive regions and allows comparative genomic analyses that facilitate our understanding of <em>Ap. gossypii</em>'s adaptation and diversification.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"176 ","pages":"Article 104215"},"PeriodicalIF":3.2,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142685596","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 unique mechanism of transfluthrin action revealed by mapping its binding sites in the mosquito sodium channel 通过绘制蚊子钠通道中的结合位点揭示高效氯氟氰菊酯的独特作用机制

IF 3.2 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2024-11-19 DOI: 10.1016/j.ibmb.2024.104214

Funmilayo Egunjobi , Felipe Andreazza , Boris S. Zhorov , Ke Dong

Pyrethroid insecticides exert their toxic action by prolonging the opening of insect voltage-gated sodium channels, resulting in the characteristic tail current during membrane repolarization in voltage clamp experiments. Permethrin (PMT) and deltamethrin (DMT), representative type I and type II pyrethroids, respectively, are predicted to bind to two lipid-exposed pyrethroid receptor sites, PyR1 and PyR2, at the lipid-exposed interfaces of repeats II/III and I/II, respectively. Transfluthrin (TF), a volatile type I pyrethroid and mosquito repellent, has received increased attention in the global combat of vector-borne human diseases. However, the electrophysiological and molecular bases of TF action on insect sodium channels remain unexplored. In this study we discovered that, unlike DMT and PMT, TF barely induces the characteristic tail current of the Aedes aegypti mosquito sodium channel (AaNa_v1-1) expressed in Xenopus oocytes. Instead, TF induces a unique persistent current. We docked TF into the AlphaFold2 model of AaNa_v1-1 and found that the tetrafluorophenyl ring of TF binds to alpha helices S5, P1, and S6, but not to the linker helices S4-S5 within either PyR1 or PyR2. In agreement with the model, functional examination of 15 AaNa_v1-1 mutants demonstrated that substitutions of DMT/PMT-sensing residues in helices S5, P1, and S6, but not in the linker-helices S4-S5, altered channel sensitivity to TF. These results revealed the unique action of TF on channel gating and suggest a distinct subtype of type I pyrethroids with a previously uncharacterized pattern of interactions with residues at the dual pyrethroid receptor sites.

拟除虫菊酯类杀虫剂通过延长昆虫电压门控钠通道的开放时间来发挥其毒性作用，从而在电压钳夹实验中产生膜再极化过程中的特征性尾流。氯菊酯（PMT）和溴氰菊酯（DMT）分别是具有代表性的 I 型和 II 型拟除虫菊酯，据推测它们会分别与位于重复序列 II/III 和 I/II 的脂质暴露界面上的两个脂质暴露的拟除虫菊酯受体位点 PyR1 和 PyR2 结合。高效氟氯氰菊酯（TF）是一种挥发性 I 型拟除虫菊酯和驱蚊剂，在全球防治病媒传播的人类疾病方面受到越来越多的关注。然而，TF 作用于昆虫钠通道的电生理学和分子基础仍有待探索。在这项研究中，我们发现，与 DMT 和 PMT 不同，TF 几乎不能诱导在爪蟾卵母细胞中表达的埃及伊蚊钠通道（AaNav1-1）的特征性尾流。相反，TF 能诱导一种独特的持续电流。我们将 TF 与 AaNav1-1 的 AlphaFold2 模型对接，发现 TF 的四氟苯基环能与 S5、P1 和 S6 α 螺旋结合，但不能与 PyR1 或 PyR2 中的 S4-S5 连接螺旋结合。与该模型一致，对 15 个 AaNav1-1 突变体进行的功能检查表明，在 S5、P1 和 S6 螺旋中的 DMT/PMT 感应残基的取代，而不是在连接螺旋 S4-S5 中的取代，会改变通道对 TF 的敏感性。这些结果揭示了 TF 对通道门控的独特作用，并表明 I 型拟除虫菊酯有一种独特的亚型，其与双拟除虫菊酯受体位点残基的相互作用模式以前从未描述过。

{"title":"A unique mechanism of transfluthrin action revealed by mapping its binding sites in the mosquito sodium channel","authors":"Funmilayo Egunjobi , Felipe Andreazza , Boris S. Zhorov , Ke Dong","doi":"10.1016/j.ibmb.2024.104214","DOIUrl":"10.1016/j.ibmb.2024.104214","url":null,"abstract":"<div><div>Pyrethroid insecticides exert their toxic action by prolonging the opening of insect voltage-gated sodium channels, resulting in the characteristic tail current during membrane repolarization in voltage clamp experiments. Permethrin (PMT) and deltamethrin (DMT), representative type I and type II pyrethroids, respectively, are predicted to bind to two lipid-exposed pyrethroid receptor sites, PyR1 and PyR2, at the lipid-exposed interfaces of repeats II/III and I/II, respectively. Transfluthrin (TF), a volatile type I pyrethroid and mosquito repellent, has received increased attention in the global combat of vector-borne human diseases. However, the electrophysiological and molecular bases of TF action on insect sodium channels remain unexplored. In this study we discovered that, unlike DMT and PMT, TF barely induces the characteristic tail current of the <em>Aedes aegypti</em> mosquito sodium channel (AaNa<sub>v</sub>1-1) expressed in <em>Xenopus</em> oocytes. Instead, TF induces a unique persistent current. We docked TF into the AlphaFold2 model of AaNa<sub>v</sub>1-1 and found that the tetrafluorophenyl ring of TF binds to alpha helices S5, P1, and S6, but not to the linker helices S4-S5 within either PyR1 or PyR2. In agreement with the model, functional examination of 15 AaNa<sub>v</sub>1-1 mutants demonstrated that substitutions of DMT/PMT-sensing residues in helices S5, P1, and S6, but not in the linker-helices S4-S5, altered channel sensitivity to TF. These results revealed the unique action of TF on channel gating and suggest a distinct subtype of type I pyrethroids with a previously uncharacterized pattern of interactions with residues at the dual pyrethroid receptor sites.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"175 ","pages":"Article 104214"},"PeriodicalIF":3.2,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142680151","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

Identification of a vital transcription factor of the alanine aminotransferase in the brown planthopper and its upstream regulatory pathways 鉴定褐飞虱丙氨酸氨基转移酶的一个重要转录因子及其上游调控途径。

IF 3.2 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2024-11-14 DOI: 10.1016/j.ibmb.2024.104212

Li-Qun Wang , Shi-Hui Li , Wen-Qing Zhang

The brown planthopper (Nilaparvata lugens) is an important insect pest of rice, and can rapidly adapt to insect-resistant rice varieties. In our previous studies, alanine aminotransferase in N. lugens (NlALT) was found to play an important role in the adaptation of the brown planthopper to resistant rice IR36. Here, we further identified CCAAT/enhancer binding protein (NlC/EBP) as a vital transcription factor of NlALT. Nlp38b in the MAPKs pathway regulated the expression of NlALT by influencing the phosphorylation level of NlC/EBP. In addition, we found that NlGRL101, a G protein-coupled receptor (GPCR), was significantly higher expressed in the N. lugens population adapted to IR36 (P-IR36). After knockdown of NlGRL101 through RNAi in P-IR36 population, lower expressions of Nlp38b and NlC/EBP, along with reduced phosphorylation levels of Nlp38b and NlC/EBP were observed; moreover, NlALT activity and honeydew amount were decreased by 15.68% and 76.08%, respectively. These results indicated that insect-resistant rice IR36 induced expression of NlGRL101, which enhanced expression of NlALT through Nlp38b and NlC/EBP. These findings are helpful for better understanding of insect adaptation to resistant crop varieties.

褐飞虱（Nilaparvata lugens）是水稻的重要害虫，能迅速适应抗虫水稻品种。我们之前的研究发现，褐飞虱体内的丙氨酸氨基转移酶（NlALT）在褐飞虱适应抗性水稻 IR36 的过程中发挥了重要作用。在此，我们进一步确定了 CCAAT/增强子结合蛋白（NlC/EBP）是 NlALT 的一个重要转录因子。MAPKs通路中的Nlp38b通过影响NlC/EBP的磷酸化水平来调控NlALT的表达。此外，我们还发现，在适应IR36（P-IR36）的N. lugens群体中，G蛋白偶联受体（GPCR）NlGRL101的表达量明显较高。在 P-IR36 群体中通过 RNAi 敲除 NlGRL101 后，观察到 Nlp38b 和 NlC/EBP 的表达量降低，Nlp38b 和 NlC/EBP 的磷酸化水平降低；此外，NlALT 活性和蜜露量分别降低了 15.68% 和 76.08%。这些结果表明，抗虫水稻IR36诱导了NlGRL101的表达，而NlGRL101通过Nlp38b和NlC/EBP增强了NlALT的表达。这些发现有助于更好地了解昆虫对抗性作物品种的适应性。

{"title":"Identification of a vital transcription factor of the alanine aminotransferase in the brown planthopper and its upstream regulatory pathways","authors":"Li-Qun Wang , Shi-Hui Li , Wen-Qing Zhang","doi":"10.1016/j.ibmb.2024.104212","DOIUrl":"10.1016/j.ibmb.2024.104212","url":null,"abstract":"<div><div>The brown planthopper (<em>Nilaparvata lugens</em>) is an important insect pest of rice, and can rapidly adapt to insect-resistant rice varieties. In our previous studies, alanine aminotransferase in <em>N. lugens</em> (<em>Nl</em>ALT) was found to play an important role in the adaptation of the brown planthopper to resistant rice IR36. Here, we further identified CCAAT/enhancer binding protein (<em>Nl</em>C/EBP) as a vital transcription factor of <em>Nl</em>ALT. <em>Nl</em>p38b in the MAPKs pathway regulated the expression of <em>Nl</em>ALT by influencing the phosphorylation level of <em>Nl</em>C/EBP. In addition, we found that <em>Nl</em>GRL101, a G protein-coupled receptor (GPCR), was significantly higher expressed in the <em>N. lugens</em> population adapted to IR36 (P-IR36). After knockdown of <em>NlGRL101</em> through RNAi in P-IR36 population, lower expressions of <em>Nlp38b</em> and <em>NlC/EBP</em>, along with reduced phosphorylation levels of <em>Nl</em>p38b and <em>Nl</em>C/EBP were observed; moreover, <em>Nl</em>ALT activity and honeydew amount were decreased by 15.68% and 76.08%, respectively. These results indicated that insect-resistant rice IR36 induced expression of <em>NlGRL101</em>, which enhanced expression of <em>Nl</em>ALT through <em>Nl</em>p38b and <em>Nl</em>C/EBP. These findings are helpful for better understanding of insect adaptation to resistant crop varieties.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"175 ","pages":"Article 104212"},"PeriodicalIF":3.2,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142638111","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

Binding properties of olfactory proteins to host volatiles, free fatty acids and cuticular hydrocarbons in the termite Reticulitermes aculabialis 白蚁嗅觉蛋白与寄主挥发物、游离脂肪酸和角质层碳氢化合物的结合特性。

IF 3.2 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2024-11-12 DOI: 10.1016/j.ibmb.2024.104211

Wenxu Yang, Chenxu Ye, Lu Wang, Jinjuan Nie, Xinyi Liu, Tiange Zhang, Wenxiu Zhang, Noor us Saba, Lingfang Yin, Lianxi Xing, Xiaohong Su

As eusocial insects prevalent in tropical and subtropical regions, termites are characterized by highly organized behaviors and exceptional adaptability, rooted in caste differentiation and chemical communication. These traits make them excellent models for studying insect social structures and ecological interactions. Investigating how termites use chemical signals to perceive and respond to their environment provides insights into their coordination and adaptation within complex ecosystems. This study delved into the chemosensory mechanisms of Reticulitermes aculabialis, examining the interactions of four olfactory proteins with 70 ligands, including host volatiles, cuticular hydrocarbons (CHCs), and free fatty acids (FFAs). Molecular docking simulations revealed varied affinities of the olfactory proteins for long-chain hydrocarbons (n-C₂₃ to n-C₂₈), suggesting a nuanced chemical communication system through specific hydrocarbon detection. RacuCSP1 and RacuCSP2 exhibited specific binding to linoleic acid and undecanoic acid, respectively, highlighting the significance of FFAs in the physiological and behavioral processes of termites. The four olfactory proteins showed a strong affinity for longifolene in fluorescence competitive binding experiments. Notably, RacuOBPs exhibited unique affinities for terpenoid volatiles such as β-lonone and neocembrene, while RacuCSPs specifically bound with terpenoids like 3-carene, myrtenol, α-pinene oxide and β-pinene indicating their critical roles in host detection. Behavioral observations following gene silencing revealed that RacuOBP5 was essential for recognizing longifolene and α-lonone recognition, while RacuCSP1 was key for detecting α-pinene in termites. These findings enhance our understanding of the termite chemosensory system and offer insights for developing precise pest management strategies.

白蚁是盛行于热带和亚热带地区的社会性昆虫，具有高度组织化的行为和超强的适应能力，其根源在于种姓分化和化学交流。这些特征使它们成为研究昆虫社会结构和生态相互作用的极佳模型。研究白蚁如何利用化学信号来感知和应对环境，可以深入了解它们在复杂生态系统中的协调和适应能力。本研究深入研究了白蚁的化学感应机制，考察了四种嗅觉蛋白与 70 种配体的相互作用，包括宿主挥发物、角质烃（CHC）和游离脂肪酸（FFA）。分子对接模拟揭示了嗅觉蛋白对长链碳氢化合物（n-C23 至 n-C28）的不同亲和力，这表明通过特异性碳氢化合物检测形成了一个微妙的化学通讯系统。RacuCSP1 和 RacuCSP2 分别表现出与亚油酸和十一烷酸的特异性结合，突显了脂肪酸在白蚁生理和行为过程中的重要性。在荧光竞争结合实验中，四种嗅觉蛋白对长芴具有很强的亲和力。值得注意的是，RacuOBPs对β-珑和新伞花烯等萜类挥发物表现出独特的亲和力，而RacuCSPs则与3-蒈烯、桃烯醇、α-氧化蒎烯和β-蒎烯等萜类化合物特异性结合，这表明它们在宿主检测中起着关键作用。基因沉默后的行为观察显示，RacuOBP5 是识别长叶烯和α-珑的关键，而 RacuCSP1 则是白蚁检测α-蒎烯的关键。这些发现加深了我们对白蚁化感系统的了解，为制定精确的害虫管理策略提供了启示。

{"title":"Binding properties of olfactory proteins to host volatiles, free fatty acids and cuticular hydrocarbons in the termite Reticulitermes aculabialis","authors":"Wenxu Yang, Chenxu Ye, Lu Wang, Jinjuan Nie, Xinyi Liu, Tiange Zhang, Wenxiu Zhang, Noor us Saba, Lingfang Yin, Lianxi Xing, Xiaohong Su","doi":"10.1016/j.ibmb.2024.104211","DOIUrl":"10.1016/j.ibmb.2024.104211","url":null,"abstract":"<div><div>As eusocial insects prevalent in tropical and subtropical regions, termites are characterized by highly organized behaviors and exceptional adaptability, rooted in caste differentiation and chemical communication. These traits make them excellent models for studying insect social structures and ecological interactions. Investigating how termites use chemical signals to perceive and respond to their environment provides insights into their coordination and adaptation within complex ecosystems. This study delved into the chemosensory mechanisms of <em>Reticulitermes aculabialis</em>, examining the interactions of four olfactory proteins with 70 ligands, including host volatiles, cuticular hydrocarbons (CHCs), and free fatty acids (FFAs). Molecular docking simulations revealed varied affinities of the olfactory proteins for long-chain hydrocarbons (n-C<sub>23</sub> to n-C<sub>28</sub>), suggesting a nuanced chemical communication system through specific hydrocarbon detection. RacuCSP1 and RacuCSP2 exhibited specific binding to linoleic acid and undecanoic acid, respectively, highlighting the significance of FFAs in the physiological and behavioral processes of termites. The four olfactory proteins showed a strong affinity for longifolene in fluorescence competitive binding experiments. Notably, RacuOBPs exhibited unique affinities for terpenoid volatiles such as β-lonone and neocembrene, while RacuCSPs specifically bound with terpenoids like 3-carene, myrtenol, α-pinene oxide and β-pinene indicating their critical roles in host detection. Behavioral observations following gene silencing revealed that RacuOBP5 was essential for recognizing longifolene and α-lonone recognition, while RacuCSP1 was key for detecting α-pinene in termites. These findings enhance our understanding of the termite chemosensory system and offer insights for developing precise pest management strategies.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"176 ","pages":"Article 104211"},"PeriodicalIF":3.2,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142610931","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

Nutrition influences immunity: Diet and host-parasite interactions 营养影响免疫力：饮食与宿主-寄生虫之间的相互作用

IF 3.2 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2024-11-06 DOI: 10.1016/j.ibmb.2024.104210

Ioannis Eleftherianos , Wei Zhang , Gianluca Tettamanti , Lillia Daley , Amr Mohamed , David Stanley

Nutrition plays a major role in host immune responses and in pathogen resistance. Understanding the network that modulates the relationship between nutrition and immunity remains a challenge. Several pathways govern the direct effects of nutrition on host immunity and the indirect effects mediated by pathogen populations. We note host microbiota also influence the intricate relationships between nutrition and immunity. The purpose of this review is to discuss recent findings from nutritional research in relation to insect immunology. We outline the relationship between diet, immunity, disease, and microbiota in insects and emphasize the significance of utilizing an integrative, multifaceted approach to grasping the influence of nutrition on immunity.

营养在宿主免疫反应和病原体抵抗中发挥着重要作用。了解调节营养与免疫之间关系的网络仍然是一项挑战。营养对宿主免疫力的直接影响和病原体种群介导的间接影响由几种途径决定。我们注意到宿主微生物群也会影响营养与免疫之间错综复杂的关系。本综述旨在讨论与昆虫免疫学有关的营养研究的最新发现。我们概述了昆虫的饮食、免疫、疾病和微生物群之间的关系，并强调了利用综合、多方面的方法来把握营养对免疫的影响的重要性。

引用次数: 0

Notch signaling regulates limb regeneration through Hes1 and HeyL in the Chinese mitten crab Notch信号通过Hes1和HeyL调控中华绒螯蟹的肢体再生

IF 3.2 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2024-10-30 DOI: 10.1016/j.ibmb.2024.104209

Ju Li , Xiaoyan Lv , Xin Zhang , Xiumei Zhao , Yuxuan Meng , Sidi Liu , Simiao Fu , Jinsheng Sun

Tissue regeneration is an efficient strategy developed by animals to compensate for damaged tissues, involving various types of progenitor cells. Deciphering the signal network that modulates the activity of these progenitors during regeneration is crucial for understanding the differences in regenerative capacities across species. In this study, we evaluated the expression profile and phenotypic function of Notch signaling during limb regeneration in arthropod Chinese mitten crabs. The expression of key components of the Notch signaling pathway was upregulated at 7-day post-autotomy (7 DPA), and declined later at 18-day post-autotomy (18 DPA). To assess the role of Notch, we injected dsRNA targeting the Notch gene into the automized area and evaluated the regeneration efficiency. Our results indicated that blocking Notch signaling led to regenerative defects, manifested by delays in the wound closure and blastema emergence processes. Furthermore, the expression of Notch target genes, Hes1 and HeyL, was significantly reduced following Notch knockdown by dsRNA. Knockdown of Hes1 specifically impaired the proliferation and expression of neural progenitor cell markers, without affecting myogenic cells. In contrast, blockage of HeyL inhibited the proliferation and expression of markers in both activated neurogenic and myogenic progenitor cells, while up-regulating markers of quiescent neural progenitor cells. These findings suggest that Notch signaling plays an important role in limb regeneration of E. sinensis by activating downstream effectors Hes1 and HeyL, regulating neurogenesis and myogenesis through distinct mechanisms.

组织再生是动物补偿受损组织的一种有效策略，涉及各种类型的祖细胞。破译再生过程中调节这些祖细胞活性的信号网络对于理解不同物种再生能力的差异至关重要。在这项研究中，我们评估了节肢动物中华绒螯蟹肢体再生过程中 Notch 信号的表达谱和表型功能。Notch信号通路关键组分的表达在断肢后7天（7 DPA）时上调，随后在断肢后18天（18 DPA）时下降。为了评估Notch的作用，我们向自动切除区域注射了靶向Notch基因的dsRNA，并评估了再生效率。结果表明，阻断Notch信号导致再生缺陷，表现为伤口闭合和胚泡出现过程的延迟。此外，dsRNA敲除Notch信号后，Notch靶基因Hes1和HeyL的表达明显减少。Hes1的敲除特异性地损害了神经祖细胞标记的增殖和表达，而不影响成肌细胞。相反，HeyL的阻断抑制了活化的神经原祖细胞和肌原祖细胞的增殖和标记表达，同时上调了静止神经祖细胞的标记。这些研究结果表明，Notch信号通过激活下游效应物Hes1和HeyL，通过不同的机制调节神经发生和肌发生，在中华鳖的肢体再生过程中发挥了重要作用。

{"title":"Notch signaling regulates limb regeneration through Hes1 and HeyL in the Chinese mitten crab","authors":"Ju Li , Xiaoyan Lv , Xin Zhang , Xiumei Zhao , Yuxuan Meng , Sidi Liu , Simiao Fu , Jinsheng Sun","doi":"10.1016/j.ibmb.2024.104209","DOIUrl":"10.1016/j.ibmb.2024.104209","url":null,"abstract":"<div><div>Tissue regeneration is an efficient strategy developed by animals to compensate for damaged tissues, involving various types of progenitor cells. Deciphering the signal network that modulates the activity of these progenitors during regeneration is crucial for understanding the differences in regenerative capacities across species. In this study, we evaluated the expression profile and phenotypic function of Notch signaling during limb regeneration in arthropod Chinese mitten crabs. The expression of key components of the Notch signaling pathway was upregulated at 7-day post-autotomy (7 DPA), and declined later at 18-day post-autotomy (18 DPA). To assess the role of Notch, we injected dsRNA targeting the Notch gene into the automized area and evaluated the regeneration efficiency. Our results indicated that blocking Notch signaling led to regenerative defects, manifested by delays in the wound closure and blastema emergence processes. Furthermore, the expression of Notch target genes, <em>Hes1</em> and <em>HeyL</em>, was significantly reduced following Notch knockdown by dsRNA. Knockdown of <em>Hes1</em> specifically impaired the proliferation and expression of neural progenitor cell markers, without affecting myogenic cells. In contrast, blockage of <em>HeyL</em> inhibited the proliferation and expression of markers in both activated neurogenic and myogenic progenitor cells, while up-regulating markers of quiescent neural progenitor cells. These findings suggest that Notch signaling plays an important role in limb regeneration of <em>E. sinensis</em> by activating downstream effectors Hes1 and HeyL, regulating neurogenesis and myogenesis through distinct mechanisms.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"175 ","pages":"Article 104209"},"PeriodicalIF":3.2,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142563592","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

Knockout of two uridine diphosphate-glycosyltransferase genes increases the susceptibility of Spodoptera litura to Bacillus thuringiensis toxins 敲除两个尿苷二磷酸-糖基转移酶基因会增加斑蝶对苏云金芽孢杆菌毒素的敏感性。

IF 3.2 2区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Insect Biochemistry and Molecular Biology

Pub Date : 2024-10-28 DOI: 10.1016/j.ibmb.2024.104208

Ensi Shao , Can Wang , Wenhui Zheng , Yige Ma , Shanshan Wang , Li Sha , Xiong Guan , Zhipeng Huang

Uridine diphosphate-glycosyltransferases (UGTs) catalyze sugar conjugation of endogenous and exogenous molecules in insects. In this study, 45 putative UGT genes in 11 families were identified from the genome of S. litura. Exposure to Bt toxins in 5th-instar larvae of the WT strain led to a significant upregulation of midgut UGT40 expression, particularly of SlUGT40D20, SlUGT40D22, and SlUGT40F25. This upregulation was not observed following exposure to chemical pesticides. Knockout of the UGT genes SlUGT40D20 and SlUGT40D22 in S. litura (mutant strains SlUGT40D20-KO and SlUGT40D22-KO) via CRISPR/Cas9-mediated mutagenesis increased susceptibility of S. litura to Bacillus thuringiensis (Bt) insecticidal proteins. However, in comparison with the wild-type (WT) strain, the mutants did not change susceptibility to chemical pesticides. Observations of 5th-instar larval midgut by electron microscopy revealed severe damage to the midgut epithelium caused by Cry1Ac toxin at 10 μg/g in the SlUGT40D20-KO strain compared to the WT. SDS-PAGE and LC MS/MS analyses identified a specific protein band corresponding to putative proteoglycans in the peritrophic matrix of the WT strain, which was absent in the SlUGT40D20-KO strain. Our study suggests an inverse correlation between expression of some UGTs and the susceptibility of S. litura larvae to some Bt toxins.

二磷酸尿苷-糖基转移酶（UGTs）可催化昆虫体内外源性分子的糖键合。本研究从 S. litura 的基因组中发现了 11 个家族中的 45 个推定 UGT 基因。WT 株系的五龄幼虫接触 Bt 毒素后，中肠 UGT40 的表达显著上调，尤其是 SlUGT40D20、SlUGT40D22 和 SlUGT40F25。在暴露于化学农药后没有观察到这种上调。通过 CRISPR/Cas9 介导的诱变，敲除 S. litura 的 UGT 基因 SlUGT40D20 和 SlUGT40D22（突变株 SlUGT40D20-KO 和 SlUGT40D22-KO），提高了 S. litura 对苏云金芽孢杆菌（Bt）杀虫蛋白的敏感性。然而，与野生型（WT）菌株相比，突变体对化学农药的敏感性没有改变。用电子显微镜观察第5龄幼虫的中肠，发现与WT相比，Cry1Ac毒素在10 μg/g浓度下对SlUGT40D20-KO株中肠上皮细胞造成了严重破坏。通过 SDS-PAGE 和 LC MS/MS 分析，在 WT 株系的营养周基质中发现了一条与推测的蛋白多糖相对应的特异性蛋白带，而 SlUGT40D20-KO 株系中则没有这种蛋白带。我们的研究表明，某些 UGTs 的表达与 S. litura 幼虫对某些 Bt 毒素的敏感性之间存在反相关性。

{"title":"Knockout of two uridine diphosphate-glycosyltransferase genes increases the susceptibility of Spodoptera litura to Bacillus thuringiensis toxins","authors":"Ensi Shao , Can Wang , Wenhui Zheng , Yige Ma , Shanshan Wang , Li Sha , Xiong Guan , Zhipeng Huang","doi":"10.1016/j.ibmb.2024.104208","DOIUrl":"10.1016/j.ibmb.2024.104208","url":null,"abstract":"<div><div>Uridine diphosphate-glycosyltransferases (UGTs) catalyze sugar conjugation of endogenous and exogenous molecules in insects. In this study, 45 putative UGT genes in 11 families were identified from the genome of <em>S. litura</em>. Exposure to Bt toxins in 5th-instar larvae of the WT strain led to a significant upregulation of midgut UGT40 expression, particularly of SlUGT40D20, SlUGT40D22, and SlUGT40F25. This upregulation was not observed following exposure to chemical pesticides. Knockout of the UGT genes <em>SlUGT40D20</em> and <em>SlUGT40D22</em> in <em>S. litura</em> (mutant strains SlUGT40D20-KO and SlUGT40D22-KO) via CRISPR/Cas9-mediated mutagenesis increased susceptibility of <em>S. litura</em> to <em>Bacillus thuringiensis</em> (Bt) insecticidal proteins. However, in comparison with the wild-type (WT) strain, the mutants did not change susceptibility to chemical pesticides. Observations of 5th-instar larval midgut by electron microscopy revealed severe damage to the midgut epithelium caused by Cry1Ac toxin at 10 μg/g in the SlUGT40D20-KO strain compared to the WT. SDS-PAGE and LC MS/MS analyses identified a specific protein band corresponding to putative proteoglycans in the peritrophic matrix of the WT strain, which was absent in the SlUGT40D20-KO strain. Our study suggests an inverse correlation between expression of some UGTs and the susceptibility of <em>S. litura</em> larvae to some Bt toxins.</div></div>","PeriodicalId":330,"journal":{"name":"Insect Biochemistry and Molecular Biology","volume":"175 ","pages":"Article 104208"},"PeriodicalIF":3.2,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142542520","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