bmo-miR-3378参与家蚕杆状病毒免疫应答调控机制的研究

In light of current information about miRNA regulation of insect immune response against viruses, especially for the mechanisms of miRNAs on immune interaction between insects and viruses, still being insufficient, and the clear fact that the expression of bmo-miR-3378, a miRNA of the silkworm, Bombyx mori, was down-regulated significantly after infected by BmNPV, systems of over-expression and inhibited-expression of bmo-miR-3378 in BmN cells are to be set up, and the cytological, pathological, and -omical effects of this miRNA to be analyzed via the integrated use of methods of molecular biology, bioinformatics and biological systematics in this project. The regulatory networks of bmo-miR-3378 in the control of expression and interaction of genes during immune response of B. mori to BmNPV will be constructed. The results of this project will contribute to understand the mechanisms of bmo-miR-3378 and its regulatory networks in inmmune interaction between B. mori immune and baculovirus, which will provide a good theoretical reference to regulation mechanism of miRNA on insect immune, new technical ideas and platform for systematic analysis of biological functions of miRNAs and their target genes, and the application foundation of early diagnosis to virus disease in silkworm, new approachs of disease defense and disease-resistant breeding, and sustainable industrial development of silkworm Bio-reactor.

针对当前有关miRNA调控昆虫免疫反应，特别是在昆虫与病毒免疫互作中的作用机制尚乏研究之现状，以重要的鳞翅目模式昆虫-家蚕为对象，在已明确bmo-miR-3378经家蚕感染BmNPV后显著下调的基础上，采用分子生物学、生物信息组学和生物系统学等技术手段，构建家蚕bmo-miR-3378过表达/抑制表达体系，分析家蚕bmo-miR-3378的细胞学效应、病理学效应和组学效应，建立bmo-miR-3378基因表达与互作调控网络，筛选并验证bmo-miR-3378靶基因及其通路相关基因的功能，以探明bmo-miR-3378及其控调网络在家蚕与BmNPV免疫互作中的作用机制，为揭示昆虫miRNA免疫调控机制提供理论参考，为miRNA及其靶基因生物功能的系统分析提供新的技术思路和分析平台，并为家蚕病毒病的早期诊断、开发防病新途径和抗病育种以及家蚕生物反应器的可持续产业化发展提供应用基础。

本项目在构建家蚕细胞bmo-miR-3378过表达/抑制表达体系的基础上，采用实时定量PCR技术、生化分析技术、蛋白组测序技术和生物信息组学等分析bmo-miR-3378表达量变化所引发的细胞生物学效应、病毒病理学效应和细胞蛋白组学效应等，筛选并验证bmo-miR-3378靶基因及其通路相关基因的功能。明确BmNPV在感染早期是通过抑制bmo-miR-3378表达来降低家蚕细胞的凋亡水平，以维持病毒的复制与增殖。杆状病毒这种通过寄主细胞小RNA来间接调控寄主细胞凋亡的反抗性机制，是通过病毒本身IAPs直接调控寄主细胞凋亡的反抗性机制的另外补充，尚属首次发现。项目结果为揭示昆虫小RNA免疫调控机制提供理论参考，为miRNA及其靶基因生物功能的系统分析提供新的技术思路和分析平台，并为家蚕病毒病的早期诊断、开发防病新途径和抗病育种以及家蚕生物反应器的可持续产业化发展提供应用基础。