玉米miR164h-5p及靶基因ZmNAC21调控丝黑穗病抗性分子机制解析

玉米丝黑穗病是由丝轴黑粉菌引起的世界性真菌土传病害，在我国北方春玉米区尤为严重。目前，在2.09区域克隆了主效抗病基因ZmWAK和ZmNL，并明确基因功能，项目组前期发现玉米miR164h-5p及负控靶基因ZmNAC21是抗丝轴黑粉菌侵染的重要调控因子，但其分子机制尚未解析。本项目以玉米miR164h-5p及靶基因ZmNAC21为研究对象，明确其组织表达及亚细胞定位，分别构建玉米miR164h-5p及靶基因的过表达和敲除载体，转化抗感玉米自交系，选取功能明确的转基因株系，结合Lnc-转录组和蛋白质组测序技术分析在转录和翻译水平调控网络；通过ChIP-seq等技术明确ZmNAC21在玉米中互作基因，同时分析转基因抗性株系与阴性对照间菌丝及玉米组织细胞形态变化，系统解析miR164h-5p负控靶基因ZmNAC21调节玉米抗丝黑穗病的分子机制，为深入阐明玉米抗丝黑穗病分子调控机理提供遗传学证据。

Maize head smut is a fungal soil-borne disease which caused by Sporisorium reilianum in the worldwide, particularly in north spring maize region. The major resistance genes, ZmWAK and ZmNL, were cloned from bin 2.09. The gene function were confirmed. In our previous study, we have found that miR164h-5p and its target gene ZmNAC21 are involved in the resistance to heat smut in maize. However, its associate dregulatory mechanism has not been reported. In this project, the tissue expression and subcellular localization will be identified. The overexpression vectors of miR164h-5p and ZmNAC21 with mutation of target site, and CRISPR vectors of miR164h-5p and ZmNAC21 will be constructed and transformed into the resistance and susceptibility maize inbred lines. The functional transgenic lines will be used to analysis through the Lnc-transcriptome and proteome. The interaction genes will be explicited with ChIP-seq and other technology. The changes of hypha and histocyte will be analyzed between transgenic resistant lines and negative control. The results obtained will help us to understand the theoretical basis underlying the molecular mechanism of maize resistance to head smut.