组合转录组和蛋白质组学分析去氢骆驼蓬碱对 Sf9 细胞的影响，揭示潜在的抗性机制。
Combined transcriptomic and proteomic analysis of harmine on Spodoptera frugiperda Sf9 cells to reveal the potential resistance mechanism.
机构信息
Key Laboratory of Natural Pesticide & Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou 510642, China; Key Laboratory of Integrated Pest Management on Crops in South China, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China.
Key Laboratory of Natural Pesticide & Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou 510642, China; Key Laboratory of Integrated Pest Management on Crops in South China, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China; Guangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology, South China Normal University, Guangzhou 510631, China.
出版信息
J Proteomics. 2020 Jan 16;211:103573. doi: 10.1016/j.jprot.2019.103573. Epub 2019 Nov 1.
The fall armyworm, Spodoptera frugiperda, is an important invasive pest and exhibits resistance to many insecticides. Harmine, a remarkable, natural β-carboline alkaloid, exhibits a variety of bioactivities and induces programmed cell death in Sf9 cells. In the present study, transcriptomic and proteomic analyses were combined to investigate the effects of harmine on Sf9 cells. A sublethal dose, 0.05 mM, was selected and the transcriptomic analysis revealed 2463 upregulated and 689 downregulated genes after harmine treatment. The most frequently enriched pathways of differentially expressed genes (DEGs) were mainly involved in drug and xenobiotic metabolism. The proteomics analysis revealed 36 upregulated and 77 downregulated proteins, and the results showed a nonlinear relationship with mRNA expression. All the genes related to detoxification and resistance in the transcriptome and DEGs were identified and annotated. Complete open reading frames of 27 cytochrome P450s (CYPs), 27 glutathione S-transferases (GSTs), 11 carboxylesterases (CarEs), 10 UDP-glucuronosyltransferases (UGTs) and 29 heat shock proteins (HSPs) were assembled and verified using qRT-PCR. Harmine exhibited a completely different detoxification mechanism from normal pesticides. The Sigma and Delta class GSTs and UGTs might play important roles, rather than CYP6 and CYP9 clans, CarEs or HSPs. BIOLOGICAL SIGNIFICANCE: Harmine, a natural β-carboline alkaloid, inhibits the cell proliferation and induces programmed cell death of Sf9 cells, which derived from Spodoptera frugiperda, an important world invasive pest. In the article, the combined transcriptomic and proteomic analysis is used to explore the potential solution for its resistance management. These results supporting that harmine can be applied as a novel adjuvant or pesticide. In addition, the systematically identified resistance-related genes in fall armyworm provide the foundation for potential resistance monitoring and management.
秋粘虫，Spodoptera frugiperda，是一种重要的入侵性害虫，对许多杀虫剂具有抗性。哈尔明，一种显著的天然β-咔啉生物碱，具有多种生物活性，并诱导 Sf9 细胞程序性细胞死亡。在本研究中，结合转录组学和蛋白质组学分析来研究哈尔明对 Sf9 细胞的影响。选择亚致死剂量 0.05 mM，转录组分析显示哈尔明处理后有 2463 个上调和 689 个下调基因。差异表达基因（DEGs）最常富集的途径主要涉及药物和外来化合物代谢。蛋白质组学分析显示 36 个上调和 77 个下调蛋白，结果与 mRNA 表达呈非线性关系。在转录组和 DEGs 中鉴定并注释了与解毒和抗性相关的所有基因。使用 qRT-PCR 组装和验证了 27 个细胞色素 P450（CYP）、27 个谷胱甘肽 S-转移酶（GST）、11 个羧酸酯酶（CarE）、10 个 UDP-葡萄糖醛酸转移酶（UGT）和 29 个热休克蛋白（HSP）的完整开放阅读框。哈尔明表现出与普通杀虫剂完全不同的解毒机制。Sigma 和 Delta 类 GST 和 UGT 可能发挥重要作用，而不是 CYP6 和 CYP9 族、CarE 或 HSPs。生物学意义：哈尔明，一种天然的β-咔啉生物碱，抑制 Sf9 细胞的增殖并诱导其程序性细胞死亡，Sf9 细胞来源于 Spodoptera frugiperda，这是一种重要的世界性入侵性害虫。在本文中，使用转录组学和蛋白质组学的组合分析来探索其抗性管理的潜在解决方案。这些结果支持将哈尔明用作新型佐剂或杀虫剂。此外，秋粘虫中系统鉴定的抗性相关基因为潜在的抗性监测和管理提供了基础。
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