甜菜夜蛾围食膜的蛋白质组鉴定
Identification of the peritrophic membrane proteome of Spodoptera exigua
梁振普；李鹏娟；王 亮；王朝兴；张小霞
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DOI：10.7679/j.issn.2095-1353.2021.031
作者单位：河南农业大学生命科学学院，郑州 450002；河南农业大学生命科学学院，郑州 450002；河南农业大学生命科学学院，郑州 450002；河南农业大学生命科学学院，郑州 450002；河南农业大学生命科学学院，郑州 450002
中文关键词：甜菜夜蛾；围食膜；液质联用技术；蛋白质组；生物信息学
英文关键词：Spodoptera exigua; peritrophic membrane; liquid chromatography and mass spectrometry; proteome; bioinformatics
中文摘要：
【目的】 甜菜夜蛾Spodoptera exigua是农业的重要害虫，为害我国蔬菜等农作物的生长。围食膜（Peritrophic membrane，PM）是昆虫体内的第一道屏障，有助于昆虫食物消化过程和保护昆虫避免细菌或病毒的侵染。本研究选取甜菜夜蛾为实验对象，通过鉴定围食膜的总蛋白成分，为探讨围食膜与病原菌相互作用的生理机制奠定基础。【方法】 选用人工饲料饲养的甜菜夜蛾5龄幼虫，剥离幼虫围食膜，利用强变性剂三氟甲磺酸（Trifluoromethane-sulfonic acid，TFMS）处理，并对围食膜蛋白进行质谱鉴定和生物信息学分析，使用BLAST鉴定蛋白的同源性，探究基因水平转移现象。【结果】 本研究共鉴定出199个围食膜蛋白。这些鉴定的蛋白可富集于84条GO term（P<0.05）；此外，所鉴定的蛋白可被注释到KEGG的73条代谢通路中，并能形成多个蛋白互作网络，分别参与能量代谢、蛋白水解、过氧化氢代谢、免疫等多种过程。同时，鉴定出与抗逆性相关蛋白，具有解毒代谢等功能，提高昆虫对农药和恶劣环境的抗性。【结论】 本研究对鉴定出的199个围食膜蛋白进行分析，初步揭示围食膜潜在的生物学功能，并提出一些害虫控制思路，为新型杀虫剂的研发提供了理论基础。
英文摘要：
[Objectives] Spodoptera exigua is a major agricultural insect pest that damages the growth of many vegetable crops. The peritrophic membrane (PM) is a semipermeable membrane secreted by insect midgut cells that plays an important role in digestion and protects insects from microbes and parasites. The aim of this study was to identify the protein constituents of the S. exigua peritrophic membrane, in order to increase understanding of the molecular interaction between the PM and pathogenic microorganisms. [Methods] The PM of 5th instar S. exigua larvae were removed and treated with trifluoromethane-sulfonic acid (TFMS). The peritrophic membrane proteome of larvae was identified by liquid chromatography and mass spectrometry (LC-MS/MS), and the results subjected to bioinformatics analysis. BLAST software was used to compare protein homology online and to explore horizontal gene transfer. [Results] A total of 199 peritrophic membrane proteins from Spodoptera exigua larvae were identified. These proteins could be enriched in 84 GO terms (P<0.05). KEGG enrichment results categorized the identified proteins into 73 metabolic pathways. The results of protein-protein interaction (PPI) analysis indicate that the identified proteins form multiple interaction networks and participate in various processes, such as energy metabolism, protein hydrolysis, hydrogen peroxide metabolism and immunity. The identified proteins included stress-related proteins which are involved in detoxification and metabolism. Such proteins can improve the resistance of insects to pesticides and harsh environmental conditions. [Conclusion] A total of 199 peritrophic membrane proteins of S. exigua larvae were identified and analyzed. The results provide useful information on the potential biological function of the PM and facilitate the development of novel, effective, biological pesticides.