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Transcriptomic and proteomic elucidation of Z chromosome dosage compensation in Helicoverpa armigera Helicoverpa armigera Z 染色体剂量补偿的转录组和蛋白质组阐释。
IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-01 DOI: 10.1111/imb.12939
Zhongyuan Deng, Yakun Zhang, Xingcheng Xie, Huihui Li, Han Guo, Xinzhi Ni, Xianchun Li

Transcriptomic data have been used to study sex chromosome dosage compensation (SCDC) in approximately 10 Lepidoptera ZW species, yielding a consensus compensation pattern of Z ZZ<AA. It remains unclear whether this compensation pattern holds when examining more Lepidoptera ZW species and/or using proteomic data to analyse SCDC. Here we combined transcriptomic and proteomic data as well as transcriptional level of six individual Z genes to reveal the SCDC pattern in Helicoverpa armigera, a polyphagous lepidopteran pest of economic importance. Transcriptomic analysis showed that the Z chromosome expression of H. armigera was balanced between male and female but substantially reduced relative to autosome expression, exhibiting an SCDC pattern of Z ZZ<AA. When using H. amigera midgut proteomic data, the SCDC pattern of this species changed from Z ZZ<AA at transcriptomic level to Z = ZZ = AA at the proteomic level. RT-qPCR analysis of transcript abundance of six Z genes found that compensation for each Z gene could vary from no compensation to overcompensation, depending on the individual genes and tissues tested. These results demonstrate for the first time the existence of a translational compensation mechanism, which is operating in addition to a translational mechanism, such as has been reported in other lepidopteran species. And the transcriptional compensation mechanism functions to accomplish Z chromosome dosage balance between the sexes (M = F on the Z chromosome), whereas the translation compensation mechanism operates to achieve dosage compensation between Z chromosome and autosome (Z = AA).

转录组数据已被用于研究约 10 个鳞翅目 ZW 种类的性染色体剂量补偿(SCDC),得出的共识补偿模式为 Z ≈ ZZ AA . $$ approx mathrm{ZZ} 。 在研究更多鳞翅目 ZW 种类和/或使用蛋白质组数据分析 SCDC 时,这种补偿模式是否成立仍不清楚。在此,我们结合转录组和蛋白质组数据以及六个 Z 基因的转录水平,揭示了 Helicoverpa armigera(一种具有重要经济价值的多食性鳞翅目害虫)的 SCDC 模式。转录组分析表明,H. armigera 的 Z 染色体表达在雌雄之间是平衡的,但相对于自体的表达大幅降低,表现出 Z ≈ ZZ AA $$ approx mathrm{ZZ} 的 SCDC 模式。当使用 H. amigera 中肠蛋白组数据时,该物种的 SCDC 模式从转录组水平的 Z ≈ ZZ AA $$ approx mathrm{ZZ} 变为蛋白组水平的 Z = ZZ = AA。对 6 个 Z 基因转录本丰度的 RT-qPCR 分析发现,每个 Z 基因的补偿可从无补偿到过度补偿不等,具体取决于测试的各个基因和组织。这些结果首次证明了翻译补偿机制的存在,这种机制是在翻译机制之外运行的,如在其他鳞翅目物种中所报道的那样。转录补偿机制的作用是实现 Z 染色体在两性之间的剂量平衡(Z 染色体上的 M = F),而翻译补偿机制的作用是实现 Z 染色体和自体之间的剂量补偿(Z = AA)。
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引用次数: 0
DNA methylation machinery is involved in development and reproduction in the viviparous pea aphid (Acyrthosiphon pisum) DNA 甲基化机制参与胎生豌豆蚜(Acyrthosiphon pisum)的发育和繁殖。
IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-26 DOI: 10.1111/imb.12936
Kane Yoon, Stephanie Williams, Elizabeth J. Duncan

Epigenetic mechanisms, such as DNA methylation, have been proposed to mediate plastic responses in insects. The pea aphid (Acyrthosiphon pisum), like the majority of extant aphids, displays cyclical parthenogenesis - the ability of mothers to switch the reproductive mode of their offspring from reproducing parthenogenetically to sexually in response to environmental cues. The pea aphid genome encodes two paralogs of the de novo DNA methyltransferase gene, dnmt3a and dnmt3x. Here we show, using phylogenetic analysis, that this gene duplication event occurred at least 150 million years ago, likely after the divergence of the lineage leading to the Aphidomorpha (phylloxerans, adelgids and true aphids) from that leading to the scale insects (Coccomorpha) and that the two paralogs are maintained in the genomes of all aphids examined. We also show that the mRNA of both dnmt3 paralogs is maternally expressed in the viviparous aphid ovary. During development both paralogs are expressed in the germ cells of embryos beginning at stage 5 and persisting throughout development. Treatment with 5-azactyidine, a chemical that generally inhibits the DNA methylation machinery, leads to defects of oocytes and early-stage embryos and causes a proportion of later stage embryos to be born dead or die soon after birth. These phenotypes suggest a role for DNA methyltransferases in reproduction, consistent with that seen in other insects. Taking the vast evolutionary history of the dnmt3 paralogs, and the localisation of their mRNAs in the ovary, we suggest there is a role for dnmt3a and/or dnmt3x in early development, and a role for DNA methylation machinery in reproduction and development of the viviparous pea aphid.

DNA 甲基化等表观遗传机制被认为是昆虫可塑性反应的中介。豌豆蚜(Acyrthosiphon pisum)与现存的大多数蚜虫一样,表现出周期性孤雌生殖--母蚜能够根据环境线索将其后代的生殖模式从孤雌生殖转换为有性生殖。豌豆蚜基因组中编码了两个新DNA甲基转移酶基因的旁系亲属,即dnmt3a和dnmt3x。在这里,我们利用系统发生学分析表明,这一基因复制事件发生在至少 1.5 亿年前,很可能是在蚜虫科(phylloxerans、adelgids 和真蚜虫)与鳞翅目昆虫科(Coccomorpha)分化之后,而且这两个对等基因保留在所有被研究蚜虫的基因组中。我们还发现,两个 dnmt3 旁系亲属的 mRNA 在胎生蚜虫的子房中都有母性表达。在发育过程中,这两个旁系亲属都在胚胎的生殖细胞中表达,从第 5 阶段开始并持续整个发育过程。5-azactyidine 是一种通常能抑制 DNA 甲基化机制的化学物质,用它处理卵母细胞和早期胚胎会导致缺陷,并导致一部分后期胚胎先天死亡或出生后不久死亡。这些表型表明 DNA 甲基转移酶在繁殖过程中的作用,这与在其他昆虫中看到的情况一致。考虑到 dnmt3 旁系亲属的漫长进化史及其 mRNA 在卵巢中的定位,我们认为 dnmt3a 和/或 dnmt3x 在早期发育中发挥作用,DNA 甲基化机制在胎生豌豆蚜的繁殖和发育中发挥作用。
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引用次数: 0
Antibacterial mechanism and structure–activity relationships of Bombyx mori cecropin A 蚕蛹素 A 的抗菌机制和结构-活性关系
IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-19 DOI: 10.1111/imb.12934
Yuyuan Tian, Hongxian Wei, Fuping Lu, Huazhou Wu, Dezhao Lou, Shuchang Wang, Tao Geng

Bombyx mori cecropin A (Bmcecropin A) has antibacterial, antiviral, anti-filamentous fungal and tumour cell inhibition activities and is considered a potential succedaneum for antibiotics. We clarified the antibacterial mechanism and structure–activity relationships and then directed the structure–activity optimization of Bmcecropin A. Firstly, we found Bmcecropin A shows a strong binding force and permeability to cell membranes like a detergent; Bmcecropin A could competitively bind to the cell membrane with the cell membrane-specific dye DiI, then damaged the membrane for the access of DiI into the cytoplasm and leading to the leakage of electrolyte and proteins. Secondly, we found Bmcopropin A could also bind to and degrade DNA; furthermore, DNA library polymerase chain reaction (PCR) results indicated that Bmcecropin A inhibited DNA replication by non-specific binding. In addition, we have identified C-terminus amidation and serine-lysine- glycine (SLG) amino acids of Bmcecropin A played critical roles in the membrane damage and DNA degradation. Based on the above results, we designed a mutant of Bmcecropin A (E9 to H, D17 to K, K33 to A), which showed higher antibacterial activity, thermostability and pH stability than ampicillin but no haemolytic activity. Finally, we speculated that Bmcecropin A damaged the cell membrane through a carpet model and drew the schematic diagram of its antibacterial mechanism, based on the antibacterial mechanism and the three-dimensional configuration. These findings yield insights into the mechanism of antimicrobial peptide–pathogen interaction and beneficial for the development of new antibiotics.

虫草素 A(Bmcecropin A)具有抗菌、抗病毒、抗丝状真菌和抑制肿瘤细胞的活性,被认为是一种潜在的抗生素继代物。首先,我们发现 Bmcecropin A 像去污剂一样对细胞膜具有很强的结合力和渗透性;Bmcecropin A 能与细胞膜特异性染料 DiI 竞争性地结合到细胞膜上,然后破坏细胞膜使 DiI 进入细胞质,导致电解质和蛋白质的泄漏。其次,我们发现Bmcopropin A还能与DNA结合并降解DNA;此外,DNA文库聚合酶链反应(PCR)结果表明,Bmcecropin A通过非特异性结合抑制DNA复制。此外,我们还发现 Bmcecropin A 的 C 端酰胺化和丝氨酸-赖氨酸-甘氨酸(SLG)氨基酸在膜损伤和 DNA 降解中起着关键作用。根据上述结果,我们设计了一种 Bmcecropin A 的突变体(E9 至 H、D17 至 K、K33 至 A),它比氨苄西林具有更高的抗菌活性、热稳定性和 pH 稳定性,但没有溶血活性。最后,我们根据抗菌机理和三维构型,通过地毯模型推测 Bmcecropin A 破坏了细胞膜,并绘制了其抗菌机理示意图。这些发现深入揭示了抗菌肽与病原体相互作用的机制,有利于新型抗生素的开发。
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引用次数: 0
The genome-wide response of Dermatophagoides pteronyssinus to cystatin A, a peptidase inhibitor from human skin, sheds light on its digestive physiology and allergenicity Dermatophagoides pteronyssinus 对胱抑素 A(一种来自人类皮肤的肽酶抑制剂)的全基因组反应揭示了其消化生理和过敏性。
IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-15 DOI: 10.1111/imb.12931
José Cristian Vidal-Quist, Félix Ortego, Stephane Rombauts, Pedro Hernández-Crespo

The digestive physiology of house dust mites (HDMs) is particularly relevant for their allergenicity since many of their allergens participate in digestion and are excreted into faecal pellets, a main source of exposure for allergic subjects. To gain insight into the mite dietary digestion, the genome of the HDM Dermatophagoides pteronyssinus was screened for genes encoding peptidases (n = 320), glycosylases (n = 77), lipases and esterases (n = 320), peptidase inhibitors (n = 65) and allergen-related proteins (n = 52). Basal gene expression and transcriptional responses of mites to dietary cystatin A, a cysteine endopeptidase inhibitor with previously shown antinutritional effect on mites, were analysed by RNAseq. The ingestion of cystatin A resulted in significant regulation of different cysteine endopeptidase and glycosylase genes. One Der p 1-like and two cathepsin B-like cysteine endopeptidase genes of high basal expression were induced, which suggests their prominent role in proteolytic digestion together with major allergen Der p 1. A number of genes putatively participating in the interaction of mites with their microbiota and acquired by horizontal gene transfer were repressed, including genes encoding the peptidase Der p 38, two 1,3-beta-glucanases, a lysozyme and a GH19 chitinase. Finally, the disruption of mite digestion resulted in the regulation of up to 17 allergen and isoallergen genes. Altogether, our results shed light on the putative role of specific genes in digestion and illustrate the connection between the digestive physiology of HDM and allergy.

屋尘螨(HDMs)的消化生理学与它们的过敏性特别相关,因为它们的许多过敏原都参与消化并排泄到粪便中,而粪便是过敏性受试者的主要接触源。为了深入了解螨虫的膳食消化情况,对 HDM Dermatophagoides pteronyssinus 的基因组进行了筛选,以寻找编码肽酶(n = 320)、糖基化酶(n = 77)、脂肪酶和酯酶(n = 320)、肽酶抑制剂(n = 65)和过敏原相关蛋白(n = 52)的基因。通过 RNAseq 分析了螨虫的基础基因表达和对膳食胱抑素 A 的转录反应,胱抑素 A 是一种半胱氨酸内肽酶抑制剂,以前曾显示对螨虫有抗营养作用。摄入胱抑素 A 对不同的半胱氨酸内肽酶和糖基化酶基因有显著的调节作用。一个类似 Der p 1 的半胱氨酸内肽酶基因和两个类似 cathepsin B 的半胱氨酸内肽酶基因的基础表达量很高,这表明它们与主要过敏原 Der p 1 一起在蛋白水解消化过程中发挥着重要作用。一些可能参与螨虫与其微生物群相互作用并通过水平基因转移获得的基因受到抑制,包括编码肽酶 Der p 38、两种 1,3-beta-葡聚糖酶、一种溶菌酶和一种 GH19 几丁质酶的基因。最后,螨虫消化的中断导致多达 17 个过敏原和异过敏原基因受到调控。总之,我们的研究结果揭示了特定基因在消化过程中的潜在作用,并说明了 HDM 消化生理与过敏之间的联系。
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引用次数: 0
A role of epigenetic mechanisms in regulating female reproductive responses to temperature in a pest beetle 表观遗传机制在调节害虫雌性生殖对温度的反应中的作用。
IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-12 DOI: 10.1111/imb.12933
Beth A. McCaw, Aoife M. Leonard, Tyler J. Stevenson, Lesley T. Lancaster

Many species are threatened by climate change and must rapidly respond to survive in changing environments. Epigenetic modifications, such as DNA methylation, can facilitate plastic responses by regulating gene expression in response to environmental cues. Understanding epigenetic responses is therefore essential for predicting species' ability to rapidly adapt in the context of global environmental change. Here, we investigated the functional significance of different methylation-associated cellular processes on temperature-dependent life history in seed beetles, Callosobruchus maculatus Fabricius 1775 (Coleoptera: Bruchidae). We assessed changes under thermal stress in (1) DNA methyltransferase (Dnmt1 and Dnmt2) expression levels, (2) genome-wide methylation and (3) reproductive performance, with (2) and (3) following treatment with 3-aminobenzamide (3AB) and zebularine (Zeb) over two generations. These drugs are well-documented to alter DNA methylation across the tree of life. We found that Dnmt1 and Dnmt2 were expressed throughout the body in males and females, but were highly expressed in females compared with males and exhibited temperature dependence. However, whole-genome methylation did not significantly vary with temperature, and only marginally or inconclusively with drug treatment. Both 3AB and Zeb led to profound temperature-dependent shifts in female reproductive life history trade-off allocation, often increasing fitness compared with control beetles. Mismatch between magnitude of treatment effects on DNA methylation versus life history effects suggest potential of 3AB and Zeb to alter reproductive trade-offs via changes in DNA repair and recycling processes, rather than or in addition to (subtle) changes in DNA methylation. Together, our results suggest that epigenetic mechanisms relating to Dnmt expression, DNA repair and recycling pathways, and possibly DNA methylation, are strongly implicated in modulating insect life history trade-offs in response to temperature change.

许多物种受到气候变化的威胁,必须迅速做出反应才能在不断变化的环境中生存。表观遗传修饰(如 DNA 甲基化)可根据环境线索调节基因表达,从而促进可塑性反应。因此,了解表观遗传反应对于预测物种在全球环境变化中的快速适应能力至关重要。在这里,我们研究了与甲基化相关的不同细胞过程对种子甲虫(Callosobruchus maculatus Fabricius 1775,鞘翅目:Bruchidae)温度依赖性生活史的功能意义。我们评估了热胁迫下(1) DNA甲基转移酶(Dnmt1和Dnmt2)表达水平、(2) 全基因组甲基化和(3) 繁殖性能的变化,其中(2)和(3)是在使用3-氨基苯甲酰胺(3AB)和斑蝥素(Zeb)处理两代后发生的。这些药物可改变整个生命树的 DNA 甲基化。我们发现,Dnmt1和Dnmt2在雄性和雌性体内均有表达,但雌性的表达量比雄性高,而且表现出温度依赖性。然而,全基因组甲基化并不随温度的变化而显著变化,也不随药物治疗的变化而显著变化。3AB和Zeb都会导致雌性生殖生活史权衡分配发生深刻的温度依赖性变化,与对照甲虫相比,往往会提高其适应性。对 DNA 甲基化的处理效果与对生活史的处理效果之间的不匹配表明,3AB 和 Zeb 有可能通过改变 DNA 修复和循环过程来改变生殖权衡,而不是 DNA 甲基化的(微妙)变化。总之,我们的研究结果表明,与 Dnmt 表达、DNA 修复和再循环途径有关的表观遗传学机制,以及可能与 DNA 甲基化有关的表观遗传学机制,与调节昆虫对温度变化的生活史权衡有很大关系。
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引用次数: 0
The RNA interference response to alphanodavirus replication in Phlebotomus papatasi sand fly cells RNA 干扰对 Phlebotomus papatasi 沙蝇细胞中阿尔汉诺达病毒复制的反应。
IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-07 DOI: 10.1111/imb.12932
Akira J. T. Alexander, Rhys H. Parry, Maxime Ratinier, Frédérick Arnaud, Alain Kohl

In this study, we identified and assembled a strain of American nodavirus (ANV) in the Phlebotomus papatasi-derived PP9ad cell line. This strain most closely resembles Flock House virus and ANV identified in the Drosophila melanogaster S2/S2R cell line. Through small RNA sequencing and analysis, we demonstrate that ANV replication in PP9ad cells is primarily targeted by the exogenous small interfering RNA (exo-siRNA) pathway, with minimal engagement from the PIWI-interacting RNA (piRNA) pathway. In mosquitoes such as Aedes and Culex, the PIWI pathway is expanded and specialised, which actively limits virus replication. This is unlike in Drosophila spp., where the piRNA pathway does not restrict viral replication. In Lutzomyia sandflies (family Psychodidae), close relatives of Phlebotomus species and Drosophila, there appears to be an absence of virus-derived piRNAs. To investigate whether this absence is due to a lack of PIWI pathway proteins, we analysed the piRNA and siRNA diversity and repertoire in PP9ad cells. Previous assemblies of P. papatasi genome (Ppap_1.0) have revealed a patchy repertoire of the siRNA and piRNA pathways. Our analysis of the updated P. papatasi genome (Ppap_2.1) has shown no PIWI protein expansion in sandflies. We found that both siRNA and piRNA pathways are transcriptionally active in PP9ad cells, with genomic mapping of small RNAs generating typical piRNA signatures. Our results suggest that the piRNA pathway may not respond to virus replication in these cells, but an antiviral response is mounted via the exo-siRNA pathway.

在这项研究中,我们在Plebotomus papatasi衍生的PP9ad细胞系中鉴定并组装了一株美洲结核病毒(ANV)。该毒株与Flock House病毒和在黑腹果蝇S2/S2R细胞系中鉴定出的ANV最为相似。通过小 RNA 测序和分析,我们证明 ANV 在 PP9ad 细胞中的复制主要是通过外源小干扰 RNA(exo-siRNA)途径进行的,PIWI-interacting RNA(piRNA)途径的参与极少。在伊蚊和库蚊等蚊子中,PIWI 途径得到了扩展和专门化,从而积极限制了病毒的复制。这与果蝇不同,果蝇的 piRNA 途径不会限制病毒复制。在 Phlebotomus 和果蝇的近亲 Lutzomyia 沙蝇(Psychodidae 科)中,似乎不存在源自病毒的 piRNA。为了研究这种缺失是否是由于缺乏 PIWI 通路蛋白所致,我们分析了 PP9ad 细胞中 piRNA 和 siRNA 的多样性和种类。先前的 P. papatasi 基因组(Ppap_1.0)组装结果显示,siRNA 和 piRNA 通路的种类繁多。我们对更新的 P. papatasi 基因组(Ppap_2.1)的分析表明,沙蝇中的 PIWI 蛋白没有扩增。我们发现,在 PP9ad 细胞中,siRNA 和 piRNA 途径都具有转录活性,小 RNA 的基因组图谱产生了典型的 piRNA 标志。我们的研究结果表明,在这些细胞中,piRNA途径可能不会对病毒复制做出反应,但会通过exo-siRNA途径做出抗病毒反应。
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引用次数: 0
Role of arylalkylamine N-acetyltransferase 7 in reproduction and limb pigmentation of Aedes aegypti 芳基烷基胺 N-乙酰转移酶 7 在埃及伊蚊的繁殖和肢体色素沉着中的作用。
IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-05-31 DOI: 10.1111/imb.12930
Yu Tang, Linlong Jiang, Yuqi Huang, Zhaohui Chen, David J. Merkler, Lei Zhang, Qian Han

Arylalkylamine N-acetyltransferase (aaNAT) is a crucial enzyme that catalyses the transfer of acetyl groups from acetyl coenzyme A to arylalkylamines and arylamines. Evolutionary studies have identified a distinct class of aaNATs specific to mosquitoes, yet their functions remain elusive. This study focuses on Ae-aaNAT7, a mosquito-unique gene in Aedes aegypti (Diptera:Culicidae), to explore its functionality. Temporal and spatial expression analysis of Ae-aaNAT7 mRNA revealed high expression during embryonic development and in first-instar larvae, with notable expression in the limbs of adult mosquitoes based on tissue expression profiling. By further employing CRISPR/Cas9 technology for loss-of-function studies, our investigation revealed a reduction in the area of white spotting in the limbs of Ae-aaNAT7 mutant adult mosquitoes. Further investigation revealed a significant decrease in the fecundity and hatchability of the mutants. Dissection of the ovaries from Ae-aaNAT7 heterozygous mutants showed a noticeable reduction in the oocyte area compared with wild type. Dissection of the exochorion of the eggs from Ae-aaNAT7 homozygous mutants consistently revealed a striking absence of mature embryos. In addition, RNA interference experiments targeting Ae-aaNAT7 in males resulted in a reduction in fecundity, but no effect on hatchability was observed. These collective insights underscore the substantial impact of Ae-aaNAT7 on reproduction and its pivotal contribution to adult limb pigmentation in Ae. aegypti. These revelations offer insights pivotal for the strategic design of future insecticide targets.

芳基烷基胺 N-乙酰转移酶(aaNAT)是一种重要的酶,可催化乙酰辅酶 A 向芳基烷基胺和芳基胺转移乙酰基。进化研究发现了蚊子特有的一类独特的 aaNATs,但它们的功能仍然难以捉摸。本研究以埃及伊蚊(Diptera:Culicidae)的一个蚊子特有基因 Ae-aaNAT7 为研究对象,探讨其功能。对 Ae-aaNAT7 mRNA 的时空表达分析表明,该基因在胚胎发育和初孵幼虫中高表达,根据组织表达谱分析,在成蚊的四肢中也有显著表达。通过进一步利用 CRISPR/Cas9 技术进行功能缺失研究,我们的调查发现 Ae-aaNAT7 突变体成蚊四肢的白斑面积有所减少。进一步调查发现,突变体的繁殖力和孵化率明显下降。对 Ae-aaNAT7 杂合突变体卵巢的解剖显示,与野生型相比,卵母细胞面积明显缩小。对Ae-aaNAT7同源突变体卵子的外胚层进行解剖后发现,成熟胚胎明显缺失。此外,针对雄性 Ae-aaNAT7 的 RNA 干扰实验导致繁殖力下降,但未观察到对孵化率的影响。这些发现都强调了Ae-aaNAT7对埃及蚁繁殖的重大影响及其对成虫肢体色素沉着的关键作用。这些发现为未来杀虫剂目标的战略设计提供了重要的启示。
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引用次数: 0
DNA methylase 1 influences temperature responses and development in the invasive pest Tuta absoluta DNA 甲基化酶 1 影响入侵害虫 Tuta absoluta 的温度反应和发育。
IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-05-29 DOI: 10.1111/imb.12919
Yanhong Tang, Huifang Zhang, Huanqing Zhu, Siyan Bi, Xiaodi Wang, Shunxia Ji, Jianhang Ji, Dongfang Ma, Cong Huang, Guifen Zhang, Nianwan Yang, Fanghao Wan, Zhichuang Lü, Wanxue Liu

DNA methylase 1 (Dnmt1) is an important regulatory factor associated with biochemical signals required for insect development. It responds to changes in the environment and triggers phenotypic plasticity. Meanwhile, Tuta absoluta Meyrick (Lepidoptera: Gelechiidae)a destructive invasive pestcan rapidly invade and adapt to different habitats; however, the role of Dnmt1 in this organism has not been elucidated. Accordingly, this study investigates the mechanism(s) underlying the rapid adaptation of Tuta absoluta to temperature stress. Potential regulatory genes were screened via RNAi (RNA interference), and the DNA methylase in Tuta absoluta was cloned by RACE (Rapid amplification of cDNA ends). TaDnmt1 was identified as a potential regulatory gene via bioinformatics; its expression was evaluated in response to temperature stress and during different development stages using real-time polymerase chain reaction. Results revealed that TaDnmt1 participates in hot/cold tolerance, temperature preference and larval development. The full-length cDNA sequence of TaDnmt1 is 3765 bp and encodes a 1254 kDa protein with typical Dnmt1 node-conserved structural features and six conserved DNA-binding active motifs. Moreover, TaDnmt1 expression is significantly altered by temperature stress treatments and within different development stages. Hence, TaDnmt1 likely contributes to temperature responses and organismal development. Furthermore, after treating with double-stranded RNA and exposing Tuta absoluta to 35°C heat shock or −12°C cold shock for 1 h, the survival rate significantly decreases; the preferred temperature is 2°C lower than that of the control group. In addition, the epidermal segments become enlarged and irregularly folded while the surface dries up. This results in a significant increase in larval mortality (57%) and a decrease in pupation (49.3%) and eclosion (50.9%) rates. Hence, TaDnmt1 contributes to temperature stress responses and temperature perception, as well as organismal growth and development, via DNA methylation regulation. These findings suggest that the rapid geographic expansion of T absoluta has been closely associated with TaDnmt1-mediated temperature tolerance. This study advances the research on ‘thermos Dnmt’ and provides a potential target for RNAi-driven regulation of Tuta absoluta.

DNA 甲基化酶 1(Dnmt1)是与昆虫发育所需的生化信号有关的重要调节因子。它能对环境变化做出反应,并引发表型可塑性。与此同时,Tuta absoluta Meyrick(鳞翅目:Gelechiidae)--一种破坏性入侵害虫--能迅速入侵并适应不同的生境;然而,Dnmt1 在该生物中的作用尚未阐明。因此,本研究调查了 Tuta absoluta 快速适应温度胁迫的机制。通过RNAi(RNA干扰)筛选了潜在的调控基因,并通过RACE(cDNA末端快速扩增)克隆了Tuta absoluta中的DNA甲基化酶。通过生物信息学确定 TaDnmt1 为潜在的调控基因;利用实时聚合酶链式反应评估了其在温度胁迫和不同发育阶段的表达情况。结果表明,TaDnmt1 参与了冷热耐受、温度偏好和幼虫发育。TaDnmt1 的全长 cDNA 序列长达 3765 bp,编码 1254 kDa 蛋白,具有典型的 Dnmt1 节点保守结构特征和六个保守的 DNA 结合活性基序。此外,TaDnmt1 的表达受温度胁迫处理和不同发育阶段的影响而发生显著变化。因此,TaDnmt1 很可能对温度反应和生物体的发育做出了贡献。此外,用双链 RNA 处理 Tuta absoluta 并将其暴露于 35°C 热休克或 -12°C 冷休克 1 小时后,其存活率会显著下降;首选温度比对照组低 2°C。此外,表皮片段变大,呈不规则折叠,表面干枯。这导致幼虫死亡率显著上升(57%),化蛹率下降(49.3%),羽化率下降(50.9%)。因此,TaDnmt1 通过 DNA 甲基化调控,有助于温度胁迫反应和温度感知,以及生物体的生长和发育。这些研究结果表明,TaDnmt1 介导的温度耐受性与 T absoluta 的快速地理扩张密切相关。这项研究推动了对 "耐温 Dnmt "的研究,并为 RNAi- 驱动的 Tuta absoluta 调控提供了潜在靶标。
{"title":"DNA methylase 1 influences temperature responses and development in the invasive pest Tuta absoluta","authors":"Yanhong Tang,&nbsp;Huifang Zhang,&nbsp;Huanqing Zhu,&nbsp;Siyan Bi,&nbsp;Xiaodi Wang,&nbsp;Shunxia Ji,&nbsp;Jianhang Ji,&nbsp;Dongfang Ma,&nbsp;Cong Huang,&nbsp;Guifen Zhang,&nbsp;Nianwan Yang,&nbsp;Fanghao Wan,&nbsp;Zhichuang Lü,&nbsp;Wanxue Liu","doi":"10.1111/imb.12919","DOIUrl":"10.1111/imb.12919","url":null,"abstract":"<p>DNA methylase 1 (<i>Dnmt1</i>) is an important regulatory factor associated with biochemical signals required for insect development. It responds to changes in the environment and triggers phenotypic plasticity. Meanwhile, <i>Tuta absoluta</i> Meyrick (Lepidoptera: Gelechiidae)<i>—</i>a destructive invasive pest<i>—</i>can rapidly invade and adapt to different habitats; however, the role of <i>Dnmt1</i> in this organism has not been elucidated. Accordingly, this study investigates the mechanism(s) underlying the rapid adaptation of <i>Tuta absoluta</i> to temperature stress. Potential regulatory genes were screened via RNAi (RNA interference), and the DNA methylase in <i>Tuta absoluta</i> was cloned by RACE (Rapid amplification of cDNA ends). <i>TaDnmt1</i> was identified as a potential regulatory gene via bioinformatics; its expression was evaluated in response to temperature stress and during different development stages using real-time polymerase chain reaction. Results revealed that <i>TaDnmt1</i> participates in hot/cold tolerance, temperature preference and larval development. The full-length cDNA sequence of <i>TaDnmt1</i> is 3765 bp and encodes a 1254 kDa protein with typical Dnmt1 node-conserved structural features and six conserved DNA-binding active motifs. Moreover, <i>TaDnmt1</i> expression is significantly altered by temperature stress treatments and within different development stages. Hence, <i>TaDnmt1</i> likely contributes to temperature responses and organismal development. Furthermore, after treating with double-stranded RNA and exposing <i>Tuta absoluta</i> to 35°C heat shock or −12°C cold shock for 1 h, the survival rate significantly decreases; the preferred temperature is 2°C lower than that of the control group. In addition, the epidermal segments become enlarged and irregularly folded while the surface dries up. This results in a significant increase in larval mortality (57%) and a decrease in pupation (49.3%) and eclosion (50.9%) rates. Hence, <i>TaDnmt1</i> contributes to temperature stress responses and temperature perception, as well as organismal growth and development, via DNA methylation regulation. These findings suggest that the rapid geographic expansion of <i>T absoluta</i> has been closely associated with <i>TaDnmt1-</i>mediated temperature tolerance. This study advances the research on ‘thermos Dnmt’ and provides a potential target for RNAi-driven regulation of <i>Tuta absoluta</i>.</p>","PeriodicalId":13526,"journal":{"name":"Insect Molecular Biology","volume":"33 5","pages":"503-515"},"PeriodicalIF":2.3,"publicationDate":"2024-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/imb.12919","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141161149","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Bombyx mori RPL13 participates in UV-induced DNA damage repair of B. mori nucleopolyhedrovirus through interaction with Bm65 Bombyx mori RPL13 通过与 Bm65 相互作用参与紫外线诱导的 B. mori nucleopolyhedrovirus DNA 损伤修复。
IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-05-27 DOI: 10.1111/imb.12928
Qi Tang, Jingjing Tang, Ceru Chen, Feifei Zhu, Qian Yu, Huiqing Chen, Liang Chen, Shangshang Ma, Keping Chen, Guohui Li

Ribosomal protein L13 (RPL13) is highly conserved in evolution. At present, the properties and functions of RPL13 have not been characterised in insects. In this study, Bombyx mori RPL13 (BmRPL13) was first found to be specifically recruited to the sites of ultraviolet (UV)-induced DNA damage and contributed to UV damage repair. Escherichia coli expressing BmRPL13 showed better resistance to UV radiation. After knocking down the expression of BmRPL13 in BmN cells, the repair speed of UV-damaged DNA slowed down. The further results showed that BmRPL13 interacted with B. mori nucleopolyhedrovirus (BmNPV) ORF65 (Bm65) protein to locate at the UV-induced DNA damage sites of BmNPV and helped repair UV-damaged viral DNA.

核糖体蛋白 L13(RPL13)在进化过程中高度保守。目前,RPL13在昆虫中的特性和功能尚未得到表征。在这项研究中,首次发现森蝽 RPL13(BmRPL13)被特异性地招募到紫外线(UV)诱导的 DNA 损伤位点,并有助于紫外线损伤的修复。表达 BmRPL13 的大肠杆菌对紫外线辐射有更好的抵抗力。在 BmN 细胞中敲除 BmRPL13 的表达后,紫外线损伤 DNA 的修复速度减慢。进一步的研究结果表明,BmRPL13与B. mori nucleopolyhedrovirus(BmNPV)ORF65(Bm65)蛋白相互作用,定位于BmNPV的紫外线诱导DNA损伤位点,帮助修复紫外线损伤的病毒DNA。
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引用次数: 0
A critical role for the nuclear protein Akirin in larval development in Henosepilachna vigintioctopunctata 核蛋白 Akirin 在 Henosepilachna vigintioctopunctata 幼虫发育过程中的关键作用。
IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-05-23 DOI: 10.1111/imb.12929
Ahmad Ali Anjum, Meng-Jiao Lin, Lin Jin, Guo-Qing Li

Akirin is a nuclear protein that controls development in vertebrates and invertebrates. The function of Akirin has not been assessed in any Coleopteran insects. We found that high levels of akirin transcripts in Henosepilachna vigintioctopunctata, a serious Coleopteran potato defoliator (hereafter Hvakirin), were present at prepupal, pupal and adult stages, especially in larval foregut and fat body. RNA interference (RNAi) targeting Hvakirin impaired larval development. The Hvakirin RNAi larvae arrested development at the final larval instar stage. They remained as stunted larvae, gradually blackened and finally died. Moreover, the remodelling of gut and fat body was inhibited in the Hvakirin depleted larvae. Two layers of cuticles, old and newly formed, were noted in the dsegfp-injected animals. In contrast, only a layer of cuticle was found in the dsakirin-injected beetles, indicating the arrest of larval development. Furthermore, the expression of three transforming growth factor-β cascade genes (Hvsmox, Hvmyo and Hvbabo), a 20-hydroxyecdysone (20E) receptor gene (HvEcR) and six 20E response genes (HvHR3, HvHR4, HvE75, HvBrC, HvE93 and Hvftz-f1) was significantly repressed, consistent with decreased 20E signalling. Conversely, the transcription of a juvenile hormone (JH) biosynthesis gene (Hvjhamt), a JH receptor gene (HvMet) and two JH response genes (HvKr-h1 and HvHairy) was greatly enhanced. Our findings suggest a critical role of Akirin in larval development in H. vigintioctopunctata.

Akirin 是一种控制脊椎动物和无脊椎动物发育的核蛋白。Akirin的功能尚未在任何鞘翅目昆虫中进行过评估。我们发现,一种严重的鞘翅目马铃薯脱叶害虫 Henosepilachna vigintioctopunctata(以下简称 Hvakirin)在蛹前期、蛹期和成虫期,尤其是在幼虫前肠和脂肪体中都存在高水平的 Akirin 转录本。针对 Hvakirin 的 RNA 干扰(RNAi)会影响幼虫的发育。Hvakirin RNAi 幼虫在幼虫末龄阶段停止发育。幼虫发育迟缓,逐渐变黑,最后死亡。此外,Hvakirin缺失幼虫的肠道和脂肪体重塑受到抑制。在注射了 dsegfp 的动物身上发现了新旧两层角质层。相比之下,注射了 dsakirin 的甲虫只有一层角质层,表明幼虫发育停止。此外,三个转化生长因子-β级联基因(Hvsmox、Hvmyo 和 Hvbabo)、一个 20-hydroxyecdysone (20E) 受体基因(HvEcR)和六个 20E 响应基因(HvHR3、HvHR4、HvE75、HvBrC、HvE93 和 Hvftz-f1)的表达受到显著抑制,这与 20E 信号的减少一致。相反,一个幼年激素(JH)生物合成基因(Hvjhamt)、一个 JH 受体基因(HvMet)和两个 JH 响应基因(HvKr-h1 和 HvHairy)的转录大大增强。我们的研究结果表明,Akirin 在 H. vigintioctopunctata 的幼虫发育过程中起着关键作用。
{"title":"A critical role for the nuclear protein Akirin in larval development in Henosepilachna vigintioctopunctata","authors":"Ahmad Ali Anjum,&nbsp;Meng-Jiao Lin,&nbsp;Lin Jin,&nbsp;Guo-Qing Li","doi":"10.1111/imb.12929","DOIUrl":"10.1111/imb.12929","url":null,"abstract":"<p>Akirin is a nuclear protein that controls development in vertebrates and invertebrates. The function of Akirin has not been assessed in any Coleopteran insects. We found that high levels of <i>akirin</i> transcripts in <i>Henosepilachna vigintioctopunctata</i>, a serious Coleopteran potato defoliator (hereafter <i>Hvakirin</i>), were present at prepupal, pupal and adult stages, especially in larval foregut and fat body. RNA interference (RNAi) targeting <i>Hvakirin</i> impaired larval development. The <i>Hvakirin</i> RNAi larvae arrested development at the final larval instar stage. They remained as stunted larvae, gradually blackened and finally died. Moreover, the remodelling of gut and fat body was inhibited in the <i>Hvakirin</i> depleted larvae. Two layers of cuticles, old and newly formed, were noted in the ds<i>egfp</i>-injected animals. In contrast, only a layer of cuticle was found in the ds<i>akirin</i>-injected beetles, indicating the arrest of larval development. Furthermore, the expression of three transforming growth factor-β cascade genes (<i>Hvsmox</i>, <i>Hvmyo</i> and <i>Hvbabo</i>), a 20-hydroxyecdysone (20E) receptor gene (<i>HvEcR</i>) and six 20E response genes (<i>HvHR3</i>, <i>HvHR4</i>, <i>HvE75</i>, <i>HvBrC</i>, <i>HvE93</i> and <i>Hvftz-f1</i>) was significantly repressed, consistent with decreased 20E signalling. Conversely, the transcription of a juvenile hormone (JH) biosynthesis gene (<i>Hvjhamt</i>), a JH receptor gene (<i>HvMet</i>) and two JH response genes (<i>HvKr-h1</i> and <i>HvHairy</i>) was greatly enhanced. Our findings suggest a critical role of Akirin in larval development in <i>H. vigintioctopunctata</i>.</p>","PeriodicalId":13526,"journal":{"name":"Insect Molecular Biology","volume":"33 6","pages":"650-661"},"PeriodicalIF":2.3,"publicationDate":"2024-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141086746","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
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Insect Molecular Biology
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