山核桃嫁接成活过程中调控生长素极性输出PIN基因的鉴定及其功能研究

PIN蛋白介导的生长素极性输出在生长素功能发挥中起关键调节作用，但相关研究主要以植物正常生长发育为出发点，且以模式植物为主要研究对象。目前，PIN蛋白介导的植物，特别是木本植物嫁接成活调控机制研究尚少见报道。本项目拟利用HPLC/GC-MS、免疫荧光和免疫胶体金技术测定不同处理、不同阶段、不同部位山核桃嫁接材料中生长素含量及定位的动态变化；基于山核桃组学测序结果克隆山核桃PIN家族基因，利用荧光定量PCR、Western Blot和原位杂交技术检测PIN基因在山核桃嫁接成活过程中的表达情况及定位，结合生长素的含量和定位，鉴定调控嫁接成活的山核桃PIN基因；同时在获得嫁接调控PIN基因及其启动子转基因株系的基础上，开展嫁接对比实验，探究PIN基因在嫁接成活过程中的调控作用。以解析PIN蛋白介导的山核桃嫁接成活过程调控机制，为促进嫁接苗在生产上的大规模应用提供参考。

Polar auxin output mediated by PIN proteins plays key regulatory roles in the function of auxin. However, related studies are taking the normal growth and development of plants as the starting point, and model plants are usually used as the main object. To date, less is known about the regulatory mechanism of PIN protein-mediated plant grafting, especially tree grafting. In the current project, the dynamic content and location changes of auxin in the grafted Chinese hickory materials with different treatment, different collecting stages and different sampling areas will be determined by HPLC/GC-MS, immunofluorescence and immune colloidal gold techniques; PIN family genes in Chinese hickory will be cloned on the basis of omics sequencing in this species. The PIN gene(s) regulating the grafting process of Chinese hickory will be identified using fluorescent quantitative PCR (qRT-PCR), Western Blot, and in situ hybridization (ISH), with the contents and locations of auxin during this process combined. Meanwhile, the mechanism of PIN gene(s) in regulating the grafting process will be investigated on the basis of identifying and grafting (with different combinations) the transgenic lines of the grafting related PIN gene(s), as well as PIN promoter(s). The aim of this project is to reveal the mechanism of PIN gene(s) in the regulation of Chinese hickory grafting process. Results from this project will not only lay a theoretical foundation for understanding the mechanism of auxin mediated grafting process regulation in Chinese hickory, but also provide practical references for facilitating the mass application of grafted seedlings in the production of Chinese hickory.