| 李怡萍1 *, 刘少凯1, 袁向群1, 郭予元1, 2.棉铃虫围食膜中Bt抗性相关蛋白的分离与鉴定[J].植物保护,2023,49(1):132-138. |
| 棉铃虫围食膜中Bt抗性相关蛋白的分离与鉴定 |
| Separation and identification of peritrophic membrane proteins related to Bt resistance in Helicoverpa armigera |
| 投稿时间：2022-07-31 修订日期：2022-08-29 |
| DOI：10.16688/j.zwbh.2022464 |
| 中文关键词: 棉铃虫 围食膜蛋白 Bt抗性 分离与鉴定 |
| 英文关键词:Helicoverpa armigera peritrophic membrane proteins Bt resistance separation and identification |
| 基金项目:国家自然科学基金(31071693，32172409); 国家重点研发计划(2017YFD0200900) |
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| 中文摘要: |
| 棉铃虫是世界性的重要农业害虫。围食膜作为昆虫抵御病原微生物入侵及有害物质的第一道天然保护性屏障, 其上可能存在与Bt抗性相关的受体蛋白。本研究以Bt Cry1Ac抗性和敏感品系的棉铃虫围食膜为对象, 采用NuPAGE电泳技术、配体杂交、质谱鉴定和生物信息学分析等技术, 测定了围食膜蛋白含量, 鉴定了蛋白质的组成及与Bt Cry1Ac毒素的结合能力。结果表明敏感品系围食膜中蛋白含量为22.19%, 抗性品系围食膜中蛋白含量为26.99%。抗、感品系棉铃虫围食膜上存在与Cry1Ac毒素结合的6个差异蛋白, 推测其中棉铃虫羧酸酯酶蛋白和血影蛋白是2个有意义的抗性相关蛋白, 2个新蛋白可能参与Bt抗性。研究证明棉铃虫围食膜上存在Bt结合蛋白且与抗性相关, 为进一步明确Bt抗性机制、制定合理的Bt抗性治理策略提供了理论依据。 |
| 英文摘要: |
| Helicoverpa armigera is an important agricultural pest worldwide. As the first natural protective barrier for insects to resist the invasion of pathogenic microorganisms and harmful substances, many receptor proteins related to Bt resistance may exist on the peritrophic membrane. In this study, we use the peritrophic membrane proteins of Cry1Ac-resistance (BtR) and susceptible strains (96S) of H. armigera as the object to analyze the content, compositions and binding ability to Cry1Ac of proteins by using NuPAGE electrophoresis, ligand blot, mass spectrometry identification and bioinformatics analysis technology. The results showed that the protein content of the peritrophic membrane of H. armigera in susceptible and resistant strains was 22.19% and 26.99%, respectively. Six peritrophic membrane proteins were found to bind with Cry1Ac by comparing the BtR and 96S strains of H. armigera . We speculated that H. armigera carboxylesterase protein and spectrin are two meaningful resistance-related proteins, and the other two proteins are suggested to be new proteins which may be related to Bt resistance. This study proves that Bt binding proteins exist in the peritrophic membrane of H. armigera and are related to Bt resistance. This study also provides a theoretical basis for clarifying Bt resistance mechanism and formulating reasonable Bt resistance management strategies. |
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