The polyphagous pest Apolygus lucorumhas become the dominant insect in Bacillus thuringiensis (Bt) cotton fields. The hormone 20-hydroxyecdysone (20E) regulates multiple events in insect development and physiology. 20E responses are controlled by pathways triggered by phospholipase C (PLC)-associated proteins. However, 20E-modulated genes whose expression is affected by PLC remain unknown. Here, isobaric tag for relative and absolute quantitation (iTRAQ) and immunoblot were carried out for comparing differentially expressed proteins (DEPs) in A. lucorumin response to 20E and the PLC inhibitor U73122, respectively. Totally 1624 DEPs were, respectively, found in the 20E/control, U73122/control, and 20E+U73122/control groups. Venn diagram analysis further revealed 8 DEPs that were shared among the three groups. Immunoblot validated these findings, which corroborated and highlighted the reliability of proteomics. KEGG enrichment analysis showed that the DEPs were included in diverse signaling pathways. The largest portion of DEPs among the three groups were categorized in metabolic pathways. In addition, DEPs among the three groups were also found to regulate the Ras-MAPK and PI3K-AKT pathways. This is the first time that iTRAQ was carried out to assess proteome alteration in A. lucorum nymphs in response to 20E and a PLC inhibitor. These findings provide novel insights into protein expression in A. lucorumin response to 20E, and a more comprehensive understanding of the function of PLC in 20E signal transduction.

中文翻译：

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基于iTRAQ的蛋白质组学谱分析，PLC介导的20E诱导蛋白在A肉中表达（Meyer-Dür）

在苏云金芽孢杆菌（Bt）棉田中，多食性害虫Apolygus lucorumhas成为主要昆虫。20-羟基蜕皮激素（20E）调节昆虫发育和生理过程中的多个事件。20E响应受磷脂酶C（PLC）相关蛋白触发的途径控制。然而，其表达受PLC影响的20E调节基因仍是未知的。在这里，进行了相对定量和绝对定量等压标记（iTRAQ）和免疫印迹，分别比较了芦荟对20E和PLC抑制剂U73122的差异表达蛋白（DEP）。在20E /对照组，U73122 /对照组和20E + U73122 /对照组中分别共发现1624个DEP。维恩图分析进一步揭示了三组之间共享的8个DEP。免疫印迹验证了这些发现，这证实并强调了蛋白质组学的可靠性。KEGG富集分析表明，DEPs包含在多种信号通路中。三组中DEP的最大部分按代谢途径分类。此外，三组中的DEPs还可以调节Ras-MAPK和PI3K-AKT途径。这是iTRAQ首次评估响应20E和PLC抑制剂的芦苇若虫蛋白质组改变。这些发现为A. lucorumin对20E的应答中的蛋白质表达提供了新颖的见解，并更全面地了解了PLC在20E信号转导中的功能。三组中DEP的最大部分按代谢途径分类。此外，三组中的DEPs还可以调节Ras-MAPK和PI3K-AKT途径。这是iTRAQ首次评估响应20E和PLC抑制剂的芦苇若虫蛋白质组改变。这些发现为A. lucorumin对20E的应答中的蛋白质表达提供了新颖的见解，并更全面地了解了PLC在20E信号转导中的功能。三组中DEP的最大部分按代谢途径分类。此外，三组中的DEPs还可以调节Ras-MAPK和PI3K-AKT途径。这是iTRAQ首次评估响应20E和PLC抑制剂的芦苇若虫蛋白质组改变。这些发现为A. lucorumin对20E的应答中的蛋白质表达提供了新颖的见解，并更全面地了解了PLC在20E信号转导中的功能。