褐飞虱类酵母共生菌响应吡虫啉胁迫的蛋白质组学解析

Brown planthopper (BPH), Nilaparvata lugens (St?l), is an important insect pest of rice production in China, which seriously threatens safety production of rice in our country. In production, chemical control is still main way of suppressing for BPH population. Appearance of resistance-insecticide of BPH resulted to outbreak of BPH population. Hence, elucidating the resistant mechanism of BPH, and exploring the delay and control way of BPH resistance will be an important subject urgent to be solved in rice production of our country. In our previous research, we had found yeast-like symbiote (YLS) in body of BPH could have important role in resistance development of BPH to imidacloprid, but molecular mechanism of the YLS in role of BPH resistance to the insecticide is still unclear. This project will use proteomics and bioinformatics methods, comprehensively analyze proteome change of resistant - and susceptible - imidacloprid YLS of BPH under the stress of imidacloprid pesticide. By comparison of proteome, mass spectrometry identification of differentially expressed proteins and analysis of gene function, key regulation proteins which relate to detoxification or degradation of insecticide will be screened. On these results, adaptation and molecular mechanism of the YLS in the insect pest to insecticide stress will be revealed. The research results will provide the theoretical foundation for reasonable explanation of resistant mechanism of BPH to neonicotinoid, and for prevention and management of BPH resistance.

褐飞虱Nilaparvata lugens (St?l)是我国水稻生产上的重要害虫，严重威胁我国水稻的安全生产。生产上，化学防治仍是控制褐飞虱种群的主要途径,但由于褐飞虱对杀虫剂产生抗药性，导致褐飞虱种群频繁大发生。因此，研究阐明褐飞虱抗药性产生发展机制、探索褐飞虱抗药性的延缓治理途径是我国水稻生产中急需解决的重要课题。前期研究表明，褐飞虱体内类酵母共生菌在褐飞虱对吡虫啉抗性发展中可能起重要作用，但尚不清楚类酵母共生菌在褐飞虱对吡虫啉产生抗药性中的分子机制。本项目拟利用蛋白质组学和生物信息学技术，全面分析褐飞虱类酵母共生菌抗吡虫啉菌株和敏感菌株在吡虫啉农药胁迫下的蛋白质组变化，通过蛋白质组的比较、差异表达蛋白的质谱鉴定及其基因功能分析，筛选与杀虫剂解毒/降解相关的关键调控蛋白，从而揭示类酵母共生菌在褐飞虱抗药性产生发展中的作用及其分子机制，为预防和治理褐飞虱的抗药性提供科学依据。

项目利用蛋白质组学和生物信息学技术，分析褐飞虱类酵母共生菌抗吡虫啉菌株和敏感菌株在吡虫啉农药胁迫下的蛋白质组表达谱差异。研究发现，敏感菌株与其500mg/L吡虫啉处理菌株相匹配的蛋白点为1063个，有150个显著性差异蛋白点。敏感菌株与抗性菌株相匹配的蛋白点为1142个，有33个显著性差异蛋白点。抗性菌株与其500mg/L吡虫啉处理菌株相匹配的蛋白点为987个，有134个显著性差异蛋白点。抗性菌株与其2000mg/L吡虫啉处理的菌株相匹配的蛋白点为1079个，有97个显著性差异蛋白点。包括共生菌菌株在吡虫啉胁迫前特异表达、胁迫后特异表达、胁迫后上调和胁迫后下调等4种差异表达模式。选取表达量差异2倍以上的蛋白点70个进行质谱鉴定，其中有55个通过质谱技术被成功鉴定，涉及到合成与代谢、细胞抗逆与防御、信号传导、转录和翻译、膜及运输、调控作用、解毒作用及未知功能蛋白等功能类别。经过分析，推测出褐飞虱共生菌中与抗药性相关的蛋白质可能有细胞色素P450、酰基辅酶A脱氢酶和细胞色素C过氧化酶等。研究结果阐明了褐飞虱类酵母共生菌在吡虫啉胁迫下的蛋白质表达谱及其变化规律，为进一步分析基因功能，筛选与杀虫剂解毒/降解相关的关键调控蛋白提供基础。