油菜根发育相关基因的克隆与功能分析

根是高等植物重要的器官，其发育状况的好坏直接影响植物体的生长状况，更影响产量。目前拟南芥根发育的研究已取得了较大的进展，但是根的发育是一个非常复杂的生物学过程，还有很多途径尚不清楚。本项目利用激活标签法获得的一个油菜大根系突变体（Big Root System, brs），通过BRS基因的克隆，结合BRS的过表达进行克隆基因的功能验证，同时应用RNAi技术和CRISPR/Cas9技术研究该基因缺失的生物学功能。对突变体根、茎中植物激素的变化进行测定，采用组织切片观察、BRS基因表达谱分析、亚细胞定位、蛋白互作研究以及BRS基因与其它根发育相关基因关系的解析，揭示BRS蛋白在油菜根发育过程中的分子机制。上述研究结果对克隆具有我国自主知识产权基因以及将该基因应用于农业生产具有重要的指导意义和参考价值。

The root is a very important organ for the higher plants, and it performs essential functions during plant development, including anchoring plants, uptaking water and nutrient, and responsing to environmental factors. Shaping root development could directly affect the status of plant growth condition, and then affect the yeild. The development of Arabidopsis root has made great progress in the past twenty years. However, the root development is a complex biological process, there are still a lot of process is not clear. In this proposal, we apply a big root system (brs) mutant obtained from activation tagging approach in Brassica napus to dissection the molecular mechanism of root development..Firstly, we will colone the BRS gene by Tail-PCR, and characterize the cloned gene by over expression of the BRS gene in wild type. In addition, we will knock down the BRS gene by RNAi interference technology or knock out BRS gene by CRISPR/Cas9 to study the biological function of BRS gene. Then we will analyze the expression profile of BRS in root, stem, leaf, flowers as well as in the histological section. The subcellular localization will determine in the transient expression of BRS-GFP fusion protein in the brassica napus cotyledon cell. The relationship between the BRS gene and other genes associated with the development of the root will be detected by Realtime-PCR. The interacted proteins with the BRS will conducted with the BIFC method in Brassica napus cotyledon cell. The study has important significance for cloning our own intellectual property rights of gene and the gene used in agricultural production.