越橘microRNA分子挖掘及越橘与杜鹃花类菌根真菌互作中的microRNA解析

运用高通量测序及生物信息学技术挖掘越橘microRNA(miRNA)，并进行基因芯片杂交验证。通过高通量测序技术分析菌根真菌不同侵染阶段未接种/接种真菌的越橘组织、接种菌根真菌带/无菌丝根组织细胞miRNA的表达差异。结合RNA酶学标记、核磁共振波谱法及生物信息学等方法，分析菌根真菌定殖相关的miRNA、pre-miRNA、miRNA－靶基因mRNA 3′UTR复合体的三级结构。探讨不同miRNA变异模式对RNA三级结构及菌根真菌定殖的影响，了解miRNA结构与其功能的相关性。利用miRNA过表达及突变等策略对miRNA功能进行分析，了解菌根真菌定殖相关miRNA及靶基因、pre-miRNA的相互作用及其对菌根真菌定殖的影响。通过筛选越橘菌根真菌定殖相关miRNA并分析其三级结构与功能相关性，可以从全新的miRNA视角深层次地剖析越橘杜鹃花类菌根真菌的定殖机理。

MicroRNAs (miRNAs) are unearthed by high-throughout sequencing and bioinformatic analysis from different tissues of Vaccinium, and validated by molecular hybridization with gene chip. The difference of miRNA expression among no-inoculated tissues and inoculated tissues of Vaccinium by ericoid mycorrhizal fungi, inoculated cells with mycelium, and inoculated cells without mycelium, are analyzed by high-throughout sequencing. Combining enzymatic probing, nuclear magnetic resonance spectroscopy with bioinformatic analysis, the tertiary structures of complex of miRNA-target gene mRNA 3′UTR, miRNA and pre-miRNA related to colonization of ericoid mycorrhizal fungi, are detemined. Via analyzing the effect on tertiary structures and the colonization of ericoid mycorrhizal fungi by different styles of miRNA variation, we will understand the relativity between the structures and function of miRNA. The function of miRNA is studied using overexpression of miRNA, mutant techniques and other methods, and the influence of mycorrhizal colonization and the interaction among miRNA, precursor and target gene related to colonization of ericoid mycorrhizal fungi are also analyzed. Via screening the miRNAs and studying the relativity between tertiary structures and function of miRNA, it is likely to understand in depth the mechanism of colonization of ericoid mycorrhizal fungi of Vaccinium via a novel perspective of miRNA.