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Establishment of diabetes mellitus model using Bombyx mori silkworms in a low-temperature environment 利用低温环境下的桑蚕建立糖尿病模型
IF 2.2 4区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-01-24 DOI: 10.1002/arch.22083
Jiaming Hou, Cheng Tan, Nan Chen, Yuan Zhou, Shaojun Huang, Huani Chen, Li Qian

Due to the high prevalence of diabetes mellitus, researchers have conducted numerous experimental animal studies. However, the mammalian diabetes model is cumbersome and expensive to operate, while the cheap and simple common silkworm diabetes model has the disadvantage of a short cycle time. Since the growth of silkworms is greatly affected by environmental factors, we extended the five-age cycle of silkworms by lowering the ambient temperature to establish a novel low-temperature silkworm diabetes model. Our goal was to determine whether the low-temperature feeding of a high-sugar diet to silkworms could serve as an effective animal model for diabetes. Also, we aimed to resolve certain issues concerning the normal temperature silkworm diabetes model, such as the short time frame for experiments and erratic fluctuations in blood sugar levels. Silkworms weighing between 0.9 and 1.0 g at the beginning of the fifth instar were selected, and we created diabetic silkworms by feeding mulberry leaves containing 4% glucose daily in a 16–20°C environment. When the silkworms were kept at a cooler temperature, the fifth instar stage lasted for an additional 9–11 days. In the model group, 83.3% of the silkworms had blood glucose levels greater than 7.8 mmol/L, while the total prevalence of diabetic silkworms was 89.8%. Moreover, JNK phosphorylation expression rose in the model group, while PI3K expression fell. Additionally, the JNK and PI3K signaling pathway expressions matched diabetic signals. Therefore, using silkworms to create a diabetes model in a cool environment is a straightforward and cost-effective approach to studying diabetes in animals.

由于糖尿病发病率高,研究人员进行了大量的动物实验研究。然而,哺乳动物糖尿病模型操作繁琐、成本高昂,而廉价、简单的普通家蚕糖尿病模型则存在周期短的缺点。由于蚕的生长受环境因素影响很大,我们通过降低环境温度延长了蚕的五龄周期,建立了一种新型的低温蚕糖尿病模型。我们的目标是确定低温饲喂高糖食物能否作为糖尿病的有效动物模型。此外,我们还希望解决常温家蚕糖尿病模型的一些问题,如实验时间短、血糖水平波动不稳定等。我们选择了体重在 0.9 至 1.0 克之间的五龄初期蚕,在 16-20°C 的环境中每天喂食含 4% 葡萄糖的桑叶,制造糖尿病蚕。当蚕在温度较低的环境中饲养时,第五龄期可多持续 9-11 天。在模型组中,83.3%的蚕血糖水平高于 7.8 mmol/L,而糖尿病蚕的总患病率为 89.8%。此外,模型组中 JNK 磷酸化表达上升,而 PI3K 表达下降。此外,JNK 和 PI3K 信号通路的表达与糖尿病信号相符。因此,利用蚕在低温环境中创建糖尿病模型是研究动物糖尿病的一种简单而经济的方法。
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引用次数: 0
Screening of insect immune suppressors using a recombinant phospholipase A2 of a lepidopteran insect 利用鳞翅目昆虫的重组磷脂酶 A2 筛选昆虫免疫抑制剂
IF 2.2 4区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-01-11 DOI: 10.1002/arch.22081
Gahyeon Jin, Yonggyun Kim

Phospholipase A2 (PLA2) catalyzes phospholipids at the sn-2 position to release free fatty acids, including arachidonic acid (AA) or its precursor. The free AA is then oxygenated into different eicosanoids, which mediate the diverse physiological processes in insects. Any inhibition of the PLA2 catalysis would give rise to serious malfunctioning in insect growth and development. An onion moth, Acrolepiopsis sapporensis, encodes four different PLA2 genes (As-PLA2AAs-PLA2D), in which As-PLA2A is dominantly expressed at all developmental stages and in different larval tissues. RNA interference of the As-PLA2A expression significantly reduced the PLA2 activity of A. sapporensis, which suffered from immunosuppression. A recombinant As-PLA2A protein was purified from a bacterial expression system, which exhibited a typical Michaelis—Menten kinetics and hence susceptible to a specific inhibitor to sPLA2 and dithiothreitol. A total of 19 bacterial metabolites derived from Xenorhabdus and Photorhabdus were screened against the recombinant As-PLA2A. Five potent metabolites were highly inhibitory and followed a competitive enzyme inhibition. These five inhibitors suppressed the immune responses of A. sapporensis by inhibiting hemocyte-spreading behavior and phenoloxidase activity. However, an addition of AA could significantly rescue the immunosuppression induced by the selected inhibitors. These studies suggest that the recombinant As-PLA2A protein can be applied for high-throughput screening of insect immunosuppressive compounds.

磷脂酶 A2(PLA2)催化磷脂的 sn-2 位释放游离脂肪酸,包括花生四烯酸(AA)或其前体。游离的 AA 随后被氧合为不同的二十烷酸,从而介导昆虫的各种生理过程。任何对 PLA2 催化作用的抑制都会导致昆虫生长和发育的严重失调。洋葱蛾(Acrolepiopsis sapporensis)编码四种不同的 PLA2 基因(As-PLA2A-As-PLA2D),其中 As-PLA2A 在所有发育阶段和不同幼虫组织中都有优势表达。对 As-PLA2A 表达的 RNA 干扰显著降低了遭受免疫抑制的 A. sapporensis 的 PLA2 活性。从细菌表达系统中纯化出的重组 As-PLA2A 蛋白表现出典型的 Michaelis-Menten 动力学,因此易受 sPLA2 和二硫苏糖醇特异性抑制剂的影响。针对重组 As-PLA2A 共筛选了 19 种来自 Xenorhabdus 和 Photorhabdus 的细菌代谢物。其中五种强效代谢物具有很强的抑制作用,并具有竞争性酶抑制作用。这五种抑制剂通过抑制血细胞扩散行为和酚氧化酶活性来抑制 A. sapporensis 的免疫反应。然而,添加 AA 可以显著缓解所选抑制剂引起的免疫抑制。这些研究表明,重组 As-PLA2A 蛋白可用于昆虫免疫抑制化合物的高通量筛选。
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引用次数: 0
Single domain von Willebrand factor type C “cytokines” and the regulation of the stress/immune response in insects 单域冯-威廉因子 C 型 "细胞因子 "与昆虫应激/免疫反应的调控
IF 2.2 4区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-01-08 DOI: 10.1002/arch.22071
Vassiliki Labropoulou, Luoluo Wang, Christiana Magkrioti, Guy Smagghe, Luc Swevers

The single domain von Willebrand factor type C (SVWC) appears in small secreted peptides that are arthropod-specific and are produced following environmental stress or pathogen exposure. Most research has focused on proteins with SVWC domain that are induced after virus infection and are hypothesized to function as “cytokines” to regulate the innate immune response. The expansion of SVWC genes in insect species indicates that many other functions remain to be discovered. Research in shrimp has elucidated the adaptability of Vago-like peptides in the innate immune response against bacteria, fungi and viruses after activation by Jak-STAT and/or Toll/Imd pathways in which they can act as pathogen-recognition receptors or cytokine-like signaling molecules. SVWC factors also appear in scorpion venoms and tick saliva, underlining their versatility to acquire new functions. This review discusses the discovery and function of SVWC peptides from insects to crustaceans and chelicerates and reveals the enormous gaps in knowledge that remain to be filled to understand this enigmatic group of secreted peptides.

单结构域的 C 型 von Willebrand 因子(SVWC)出现在小型分泌肽中,这些肽具有节肢动物特异性,在环境压力或接触病原体后产生。大多数研究都集中在病毒感染后诱导的具有 SVWC 结构域的蛋白质上,这些蛋白质被认为具有 "细胞因子 "的功能,可以调节先天性免疫反应。昆虫物种中 SVWC 基因的扩展表明,还有许多其他功能有待发现。对虾的研究已经阐明了Vago-like肽在先天性免疫反应中的适应性,在被Jak-STAT和/或Toll/Imd途径激活后,它们可以作为病原体识别受体或细胞因子样信号分子来对抗细菌、真菌和病毒。SVWC 因子还出现在蝎子毒液和蜱唾液中,凸显了它们获得新功能的多功能性。这篇综述讨论了从昆虫到甲壳类动物和螯足类动物中 SVWC 肽的发现和功能,并揭示了要了解这组神秘的分泌肽,还有大量知识空白有待填补。
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引用次数: 0
K64 acetylation of heat shock protein 90 suppresses nucleopolyhedrovirus replication in Bombyx mori 热休克蛋白 90 的 K64 乙酰化可抑制核多角体病毒在桑蚕体内的复制
IF 2.2 4区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-01-05 DOI: 10.1002/arch.22079
Xizhen Zhang, Shiyi Ma, Chaoguang Gu, Miao Hu, Meng Miao, Yanping Quan, Wei Yu

HSP90 is a highly conserved chaperone that facilitates the proliferation of many viruses, including silkworm (bombyx mori) nucleopolyhedrovirus (BmNPV), but the underlying regulatory mechanism was unclear. We found that suppression of HSP90 by 17-AAG, a HSP90-specific inhibitor, significantly reduced the expression of BmNPV capsid protein gp64 and viral genome replication, whereas overexpression of B. mori HSP90(BmHSP90) promoted BmNPV replication. Furthermore, in a recent study of the lysine acetylome of B. mori infected with BmNPV, we focused on the reduced viral proliferation due to changes of BmHSP90 lysine acetylation. Site-directed introduction of acetylated (K/Q) or deacetylated (K/R) mimic mutations into BmHSP90 revealed that lysine 64 (K64) acetylation activated the JAK/STAT pathway and reduced BmHSP90 ATPase activity, leading to diminished chaperone activity and ultimately inhibiting BmNPV proliferation. In this study, a single lysine 64 acetylation change of BmHSP90 was elucidated as a model of posttranslational modifications occurring in the wake of host-virus interactions, providing novel insights into potential antiviral strategies.

HSP90是一种高度保守的伴侣蛋白,它能促进包括家蚕(bombyx mori)核多角体病毒(BmNPV)在内的许多病毒的增殖,但其潜在的调控机制尚不清楚。我们发现,HSP90特异性抑制剂17-AAG抑制HSP90可显著降低BmNPV囊膜蛋白gp64的表达和病毒基因组的复制,而过表达B. mori HSP90(BmHSP90)则可促进BmNPV的复制。此外,在最近对感染了 BmNPV 的 B. mori 的赖氨酸乙酰化组的研究中,我们重点关注了由于 BmHSP90 赖氨酸乙酰化的变化而导致的病毒增殖的减少。将乙酰化(K/Q)或去乙酰化(K/R)模拟突变定点导入 BmHSP90 发现,赖氨酸 64(K64)乙酰化激活了 JAK/STAT 通路,降低了 BmHSP90 ATPase 活性,导致伴侣蛋白活性降低,最终抑制了 BmNPV 的增殖。本研究阐明了 BmHSP90 单个赖氨酸 64 乙酰化变化,将其作为宿主与病毒相互作用后发生的翻译后修饰的模型,为潜在的抗病毒策略提供了新的见解。
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引用次数: 0
Different development and fecundity between Spodoptera frugiperda USA and China populations, influenced by ecdysone-related genes 受蜕皮激素相关基因影响,美国和中国种群的鞘翅目昆虫发育和繁殖力不同
IF 2.2 4区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-01-05 DOI: 10.1002/arch.22074
Du-Yeol Choi, Hyoung-ho Mo, Youngjin Park

The fall armyworm (FAW), Spodoptera frugiperda, is one of the most harmful plant pests in the world and is globally distributed from the American continent to the Asian region. The FAW USA population (Sf-USA) and China population (Sf-CHN), which belong to corn strain, showed different developmental periods and fecundity rates in lab conditions. Sf-USA had faster development and higher fecundity compared with Sf-CHN. To examine these differences, transcriptomic data from two FAW populations were analyzed and compared. Twelve gigabytes of transcripts were read from each sample and 21,258 differentially expressed genes (DEGs) were detected. DEGs with log2 fold change ≥ 2 were identified and compared in two populations. In comparison to the Sf-CHN, we discovered that 3471 and 3851 individual DEGs upregulated and downregulated, respectively. Comparing transcriptome profiles for differential gene expression revealed several DEGs, including 39 of ecdysone (E)-, 25 of juvenile hormone-, and 15 of insulin-related genes. We selected six of E-related genes, such as Neverland, Shade, Ecdysone receptor, Ecdysone-inducible protein 74 (E74), E75, and E78 from DEGs. Gene expressions were suppressed by RNA interference to confirm the physiological functions of the selected genes from Sf-USA. The Sf-USA showed developmental retardation and a decrease in fecundity rate by suppression of E-related genes. These findings show that biological characteristics between Sf-USA and Sf-CHN are influenced by E-related genes.

秋虫(Spodoptera frugiperda)是世界上最有害的植物害虫之一,从美洲大陆到亚洲地区均有分布。属于玉米品系的FAW美国种群(Sf-USA)和中国种群(Sf-CHN)在实验室条件下表现出不同的发育期和繁殖率。与 Sf-CHN 相比,Sf-USA 发育更快,繁殖率更高。为了研究这些差异,我们分析并比较了两个草履虫种群的转录组数据。从每个样本中读取了 12 千兆字节的转录本,检测到 21,258 个差异表达基因(DEG)。对两个种群中对折变化对数≥ 2 的 DEGs 进行了鉴定和比较。与 Sf-CHN 相比,我们发现分别有 3471 和 3851 个 DEG 上调和下调。比较转录组图谱的差异基因表达发现了几个 DEGs,包括 39 个蜕皮激素(E)相关基因、25 个幼年激素相关基因和 15 个胰岛素相关基因。我们从 DEGs 中选择了 6 个与蜕皮激素相关的基因,如 Neverland、Shade、蜕皮激素受体、蜕皮激素诱导蛋白 74 (E74)、E75 和 E78。通过RNA干扰抑制基因表达,证实了Sf-USA所选基因的生理功能。通过抑制E相关基因,Sf-USA表现出发育迟缓和繁殖率下降。这些发现表明,Sf-USA和Sf-CHN的生物学特性受到E相关基因的影响。
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引用次数: 0
DNA barcoding of the genus Gampsocleis (Orthoptera, Tettigoniidae) from China 中国 Gampsocleis 属(直翅目,Tettigoniidae)的 DNA 条形编码
IF 2.2 4区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-01-05 DOI: 10.1002/arch.22070
Pu Zhang, Yuting Cai, Lan Ma, Jinyan Chai, Zhijun Zhou

DNA barcoding is a useful addition to the traditional morphology-based taxonomy. A ca. 650 bp fragment of the 5′ end of mitochondrial cytochrome c oxidase subunit I (hereafter COI-5P) DNA barcoding was sued as a practical tool for Gampsocleis species identification. DNA barcodes from 889 specimens belonging to 8 putative Gampsocleis species was analyzed, including 687 newly generated DNA barcodes. These barcode sequences were clustered/grouped into Operational Taxonomic Units (OTUs) using the criteria of five algorithms, namely Barcode Index Number (BIN) System, Assemble Species by Automatic Partitioning (ASAP), a Java program uses an explicit, determinate algorithm to define Molecular Operational Taxonomic Unit (jMOTU), Generalized Mixed Yule Coalescent (GMYC), and Bayesian implementation of the Poisson Tree Processes model (bPTP). The Taxon ID Tree grouped sequences of morphospecies and almost all MOTUs in distinct nonoverlapping clusters. Both long- and short-winged Gampsocleis species are reciprocally monophyletic in the Taxon ID Tree. In BOLD, 889 barcode sequences are assigned to 17 BINs. The algorithms ASAP, jMOTU, bPTP and GMYC clustered the barcode sequences into 6, 13, 10, and 23 MOTUs, respectively. BIN, ASAP, and bPTP algorithm placed three long-winged species, G. sedakovii, G. sinensis and G. ussuriensis within the same MOTU. All species delimitation algorithms split two short-winged species,G. fletcheri and G. gratiosa into at least two MOTUs each, except for ASAP algorithm. More detailed molecular and morphological integrative studies are required to clarify the status of these MOTUs in the future.

DNA 条形码是对传统形态分类法的有益补充。线粒体细胞色素 c 氧化酶亚单位 I(以下简称 COI-5P)5′端的约 650 bp DNA 条形码片段被认为是鉴定 Gampsocleis 物种的实用工具。分析了来自 8 个推定 Gampsocleis 种类的 889 个标本的 DNA 条形码,其中包括 687 个新生成的 DNA 条形码。这些条形码序列按照五种算法的标准聚类/分组为操作分类单元(OTUs),这五种算法分别是条形码索引号系统(BIN)、通过自动分区组装物种(ASAP)、使用明确、确定算法定义分子操作分类单元的 Java 程序(jMOTU)、广义混合尤勒聚合(GMYC)和泊松树过程模型的贝叶斯实现(bPTP)。分类群 ID 树将形态种的序列和几乎所有的 MOTUs 分成不同的非重叠群。在分类群 ID 树中,长翅和短翅的 Gampsocleis 物种互为单系。在 BOLD 中,889 个条形码序列被分配到 17 个 BIN 中。ASAP、jMOTU、bPTP 和 GMYC 算法分别将条形码序列聚类为 6、13、10 和 23 个 MOTU。BIN、ASAP 和 bPTP 算法将三个长翅种(G. sedakovii、G. sinensis 和 G. ussuriensis)归入同一 MOTU。除 ASAP 算法外,所有物种划分算法都将两个短翅种(G. fletcheri 和 G. gratiosa)划分到至少两个 MOTU 中。未来需要进行更详细的分子和形态学综合研究,以明确这些MOTU的地位。
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引用次数: 0
Ecdysone and gene expressions for chromatin remodeling, histone modification, and Broad Complex in relation to pupal commitment in Bombyx mori 蜕皮激素与染色质重塑、组蛋白修饰和 Broad Complex 的基因表达,以及与森双翅目昆虫蛹的形成有关的关系
IF 2.2 4区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-01-02 DOI: 10.1002/arch.22076
Maki Saito, Shota Fujimoto, Hideki Kawasaki

In the present study, we tried to clarify when and how pupal commitment (PT) better to use PC occurs and what is involved in the PT of Bombyx mori. To clarify this, we examined the responsiveness of a wing disc to ecdysone, referring to metamorphosis-related BR-C, development-related Myc and Wnt, and chromatin remodeling-related genes at around the predicted PT stage of the Bombyx wing disc. Wing disc responsiveness to juvenile hormone (JH) and ecdysone was examined using Methoprene and 20-hydroxyecdysone (20E) in vitro. The body weight of B. mori increased after the last larval ecdysis, peaked at Day 5 of the fifth larval instar (D5L5), and then decreased. The responsiveness of the wing disc to JH decreased after the last larval ecdysis up to D3L5. Bmbr-c (the Broad Complex of B. mori) showed enhanced expression in D4L5 wing discs with 20E treatment. Some chromatin remodeler and histone modifier genes (Bmsnr1, Bmutx, and Bmtip60) showed upregulation after being cultured with 20E in D4L5 wing discs. A low concentration of 20E is suggested to induce responsiveness to 20E in D4L5 wing discs. Bmbr-c, Bmsnr1, Bmutx, and Bmtip60 were upregulated after being cultured with a low concentration of 20E in D4L5 wing discs. The expression of Bmmyc and Bmwnt1 did not show a change after being cultured with or without 20E in D4L5 wing discs, while enhanced expression was observed with 20E in D5L5 wing discs. From the present results, we concluded that PT of the wing disc of B. mori occurred beginning on D4L5 with the secretion of low concentrations of ecdysteroids. Bmsnr1, Bmutx, Bmtip60, and BR-C are also involved.

在本研究中,我们试图弄清更好地利用 PC 的蛹期承诺(PT)是何时和如何发生的,以及什么参与了森双翅目的蛹期承诺。为了弄清这一点,我们研究了在预计的双翅目翼盘PT阶段左右,翼盘对蜕皮激素的反应性,以及与变态相关的BR-C、与发育相关的Myc和Wnt以及与染色质重塑相关的基因。在体外使用甲氧苄啶和 20-羟基蜕皮激素(20E)检测了翅盘对幼年激素(JH)和蜕皮激素的反应。森蝇的体重在最后一次幼虫蜕皮后增加,在第五龄幼虫(D5L5)的第5天达到峰值,然后下降。翅盘对 JH 的反应性在最后一次幼虫蜕皮后至 D3L5 期下降。20E处理后,Bmbr-c(B. mori的Broad复合体)在D4L5翅盘中的表达增强。一些染色质重塑基因和组蛋白修饰基因(Bmsnr1、Bmutx 和 Bmtip60)在 D4L5 翅盘中经 20E 培养后出现上调。低浓度的 20E 可诱导 D4L5 翼片对 20E 产生反应。用低浓度 20E 培养 D4L5 翼片后,Bmbr-c、Bmsnr1、Bmutx 和 Bmtip60 表达上调。Bmmyc和Bmwnt1的表达在D4L5翼盘中用或不用20E培养后没有变化,而在D5L5翼盘中用20E培养后表达增强。根据本研究结果,我们得出结论:从 D4L5 开始,随着低浓度蜕皮激素的分泌,B. mori 的翼盘开始出现 PT。Bmsnr1、Bmutx、Bmtip60 和 BR-C 也参与其中。
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引用次数: 0
Limits of mitochondrial genes in delimiting species within a Carbula species complex (Hemiptera: Pentatomidae) 线粒体基因在划分 Carbula 种群(半翅目:五蠹科)内物种方面的局限性
IF 2.2 4区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2023-12-26 DOI: 10.1002/arch.22075
Jiayue Zhou, Shujing Wang, Shengjie Yu, Yanfei Li, Mu Qiao, Qing Zhao, Edward Hughes, Huaxi Liu, Wenjun Bu

Molecular data has become a powerful tool for species delimitation, particularly among those that present limited morphological differences; while the mitochondrial genome, with its moderate length, low cost of sequencing and fast lineage sorting, has emerged as a practical data set. Due to the limited morphological differences among the closely related species of Carbula Stål 1865, the species boundaries between Carbula abbreviata (Motschulsky, 1866), Carbula humerigera (Uhler, 1860), and Carbula putoni (Jakovlev, 1876) have remained particularly unclear. In this study, we applied two phylogenetic reconstruction methods to two data sets (mitogenome and COI) to assess the phylogeny of Carbula distributed in Asia, and five species delimitation methods to determine the boundaries between East Asian Carbula species. Our phylogenetic analyses showed Carbula to be paraphyletic; the seven known species distributed within East Asia to form a single monophyletic group, and within this, C. abbreviata, C. humerigera, C. putoni and middle-type to comprise a C. humerigera species complex. Our results show that mitogenome data alone, while effective in the differentiation of more distantly related Carbula species, is not sufficient to accurately delimit the species within this newly described complex.

分子数据已成为物种划分的有力工具,尤其是在形态差异有限的物种中;而线粒体基因组长度适中、测序成本低、世系划分快,已成为实用的数据集。由于近缘种 Carbula Stål 1865 的形态差异有限,Carbula abbreviata(Motschulsky,1866 年)、Carbula humerigera(Uhler,1860 年)和 Carbula putoni(Jakovlev,1876 年)之间的种界线一直特别不清楚。在本研究中,我们对两个数据集(有丝分裂基因组和COI)采用了两种系统发育重建方法来评估分布在亚洲的鲤形目系统发育,并采用了五种物种划分方法来确定东亚鲤形目物种之间的界限。我们的系统发育分析表明,胭脂鱼属于副系;分布在东亚的7个已知物种组成一个单系群,其中C. abbreviata、C. humerigera、C. putoni和中间型组成C. humerigera物种群。我们的研究结果表明,单靠有丝分裂基因组数据虽然能有效区分亲缘关系较远的鲫鱼物种,但不足以准确划分这个新描述的复合群中的物种。
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引用次数: 0
Identification and characteristic analysis of an extracellular signal-regulated kinase from Ostrinia furnacalis Guenée 来自 Ostrinia furnacalis Guenée 的细胞外信号调节激酶的鉴定和特征分析
IF 2.2 4区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2023-12-26 DOI: 10.1002/arch.22077
Shuzhong Li, Fuqiang Xu, Yiqiang Zhang, Zupeng Gao, Zhaoyang Han, Congjing Feng

The extracellular signal-regulated kinase (ERK) pathway, a critical genetic determinant, controls diverse physiological functions, including innate immunity, development, and stress response. In the current study, a full-length cDNA (1592bp) encoding the ERK gene (OfERK) was cloned from Ostrinia furnacalis Guenée (GenBank accession number: MF797866). The open reading frame of the OfERK gene encoded 364 amino acids and shared 96.43%–98.08% amino acid identities with other insect mitogen-activated protein kinases. For spatiotemporal analysis of the expression pattern, OfERK exhibited a significant peak expression on the 3rd day of the pupa stage and showed the highest expression in hemocytes specifically. Indirect immunofluorescence assays and immuno-electron microscopy revealed a wide distribution of the OfERK protein in hemocytes and epidermis. Moreover, the results demonstrated that the Bt Cry1Ab-activated toxin significantly induces the expression of OfERK. Other genes related to immune response, development, and stress response exhibited dynamic changes in expression after Cry1Ab oral treatment. The expression of OfERK was downregulated through RNA interference, and the correlation of its expression with other related genes was verified using quantitative real-time polymerase chain reaction. Our study provides valuable insights into the regulatory mechanism of ERK in insects for future studies.

细胞外信号调节激酶(ERK)通路是一个关键的遗传决定因素,控制着多种生理功能,包括先天免疫、发育和应激反应。本研究从 Ostrinia furnacalis Guenée(GenBank登录号:MF797866)中克隆了编码 ERK 基因(OfERK)的全长 cDNA(1592bp)。OfERK 基因的开放阅读框编码 364 个氨基酸,与其他昆虫的丝裂原活化蛋白激酶有 96.43%-98.08% 的氨基酸相同性。在表达模式的时空分析中,OfERK 在蛹期的第 3 天表现出明显的表达峰值,尤其在血细胞中的表达量最高。间接免疫荧光测定和免疫电镜显示,OfERK 蛋白在血细胞和表皮中分布广泛。此外,研究结果表明,Bt Cry1Ab激活的毒素能显著诱导OfERK的表达。其他与免疫反应、发育和应激反应相关的基因在口服 Cry1Ab 后的表达也发生了动态变化。通过 RNA 干扰下调了 OfERK 的表达,并利用实时定量聚合酶链反应验证了其与其他相关基因表达的相关性。我们的研究为今后研究ERK在昆虫体内的调控机制提供了有价值的见解。
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引用次数: 0
Fitness effect and transcription profile reveal sublethal effect of nitenpyram on the predator Chrysopa pallens (Neuroptera: Chrysopidae) 硝虫酰胺对捕食者 Chrysopa pallens(神经鞘翅目:鞘翅目)的亚致死效应及其转录特征
IF 2.2 4区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2023-12-19 DOI: 10.1002/arch.22073
Qiankun Du, Yongpan Shan, Hongyan Hu, Changcai Wu, Dan Wang, Xianpeng Song, Yajie Ma, Jianping Xi, Xiangliang Ren, Xiaoyan Ma, Yan Ma

Although neonicotinoids are widely used and important insecticide, there are growing concerns about their effect on nontarget insects and other organisms. Moreover, the effects of nitenpyram (NIT), a second generation of neonicotinoid insecticides, on Chrysopa pallens are still unclear. Therefore, this study purposed to investigate the acute toxicity of NIT to C. pallens using the spotting method. To examine the potential effects of a sublethal dose of NIT (LD30, 1.85 ng of active ingredient per insect) on C. pallens, we constructed the life tables and analyzed the transcriptome data. The life table results showed that the period of second instar larvae, adult pre-oviposition period and total pre-oviposition period were significantly prolonged after exposure to sublethal dose of NIT, but had no significant effects on the other instars, longevity, oviposition days, and fecundity. The population parameters, including the preadult survival rate, gross reproduction rate, net reproductive rate, the intrinsic rate of increase, and finite rate of increase, were not significantly affected, and only the mean generation time was significantly prolonged by NIT. Transcriptome analysis showed that there were 68 differentially expressed genes (DEGs), including 50 upregulated genes and 18 downregulated genes. Moreover, 13 DEGs related to heat shock protein, nose resistant to fluoxetine protein 6, and prophenoloxidas were upregulated. This study showed the potential effects of sublethal doses of NIT on C. pallens and provided a theoretical reference for the comprehensive application of chemical and biological control in integrated pest management.

尽管新烟碱类杀虫剂是一种广泛使用的重要杀虫剂,但人们越来越关注其对非目标昆虫和其他生物的影响。此外,第二代新烟碱类杀虫剂硝虫吡喃(NIT)对苍蝇的影响仍不清楚。因此,本研究采用斑点法研究 NIT 对苍术的急性毒性。为了研究亚致死剂量 NIT(LD30,每只昆虫 1.85 纳克有效成分)对苍蝇的潜在影响,我们构建了生命表并分析了转录组数据。生命表结果表明,暴露于亚致死剂量的NIT后,二龄幼虫期、成虫产卵前期和总产卵前期显著延长,但对其他龄期、寿命、产卵天数和繁殖力无显著影响。种群参数,包括成虫前存活率、总繁殖率、净繁殖率、内在增长率和有限增长率均未受到显著影响,只有平均世代时间受到 NIT 的影响而显著延长。转录组分析表明,有68个差异表达基因(DEGs),包括50个上调基因和18个下调基因。此外,13 个与热休克蛋白、鼻抗氟西汀蛋白 6 和丙酰氧胺相关的 DEGs 上调。该研究显示了亚致死剂量NIT对苍蝇的潜在影响,为害虫综合防治中化学防治和生物防治的综合应用提供了理论参考。
{"title":"Fitness effect and transcription profile reveal sublethal effect of nitenpyram on the predator Chrysopa pallens (Neuroptera: Chrysopidae)","authors":"Qiankun Du,&nbsp;Yongpan Shan,&nbsp;Hongyan Hu,&nbsp;Changcai Wu,&nbsp;Dan Wang,&nbsp;Xianpeng Song,&nbsp;Yajie Ma,&nbsp;Jianping Xi,&nbsp;Xiangliang Ren,&nbsp;Xiaoyan Ma,&nbsp;Yan Ma","doi":"10.1002/arch.22073","DOIUrl":"https://doi.org/10.1002/arch.22073","url":null,"abstract":"<p>Although neonicotinoids are widely used and important insecticide, there are growing concerns about their effect on nontarget insects and other organisms. Moreover, the effects of nitenpyram (NIT), a second generation of neonicotinoid insecticides, on <i>Chrysopa pallens</i> are still unclear. Therefore, this study purposed to investigate the acute toxicity of NIT to <i>C. pallens</i> using the spotting method. To examine the potential effects of a sublethal dose of NIT (LD<sub>30</sub>, 1.85 ng of active ingredient per insect) on <i>C. pallens</i>, we constructed the life tables and analyzed the transcriptome data. The life table results showed that the period of second instar larvae, adult pre-oviposition period and total pre-oviposition period were significantly prolonged after exposure to sublethal dose of NIT, but had no significant effects on the other instars, longevity, oviposition days, and fecundity. The population parameters, including the preadult survival rate, gross reproduction rate, net reproductive rate, the intrinsic rate of increase, and finite rate of increase, were not significantly affected, and only the mean generation time was significantly prolonged by NIT. Transcriptome analysis showed that there were 68 differentially expressed genes (DEGs), including 50 upregulated genes and 18 downregulated genes. Moreover, 13 DEGs related to heat shock protein, nose resistant to fluoxetine protein 6, and prophenoloxidas were upregulated. This study showed the potential effects of sublethal doses of NIT on <i>C. pallens</i> and provided a theoretical reference for the comprehensive application of chemical and biological control in integrated pest management.</p>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138739816","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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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Archives of Insect Biochemistry and Physiology
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