MicroRNA对二化螟免疫信号通路p38 MAPK的调控功能分析

MicroRNA（miRNA）参与调控生物的许多重要生理过程，前人研究显示一些昆虫miRNAs的靶基因是免疫信号通路MAPK中的关键因子。申请人前期研究发现二化螟p38 MAPK路径在Cry1Ca诱导的细胞免疫反应中起关键作用，但二化螟miRNAs对细胞防御信号通路p38 MAPK的调控机制仍不清晰。因此本项目拟通过构建二化螟对Cry1Ca防御相关的miRNAs文库和分析，筛选差异表达miRNAs，通过预测靶基因，分析miRNAs对p38 MAPK信号通路的调控功能，然后通过双荧光素报告基因系统体外验证miRNAs的靶标互作关系，通过qRT-PCR和RNA免疫共沉淀试验体内验证miRNAs对p38 MAPK信号通路的调控功能。研究结果不仅对深入理解二化螟对Bt毒素的防御机制具有重要意义，同时为提高Cry毒素对靶标昆虫的毒力、延缓靶标昆虫对转Bt抗虫作物的抗性发展提供新的思路和依据。

The microRNA (miRNA) was proved to be involved in regulation of many biological pathways. There are a few insect miRNAs that was known to target genes in MAPK pathway. Our previous data proved that the C. suppressalis p38 MAPK is involved in the cellular defense against Cry1Ca toxin. Based on above results, the present program would focus on the involvement of miRNA in cellular defense against Cry1Ca toxin. The high-throughput sequencing, target gene prediction, dual-luciferase reporter gene assay and co-Immunoprecipitation system would be conducted to elucidate the function of miRNA in cellular defense regulation. Our results would improve the understanding of the insect defense mechanism against Cry toins, and would also benefit improving Cry toxin toxicity and delaying resistance development of target insects.