红皮梨是目前市场上极具吸引力和发展潜力的果品类型，其果皮红色主要由花青苷的合成与积累所致。项目组前期研究发现花青苷合成的转录调控因子是影响梨果皮着色的重要原因，并利用cDNA-AFLP 以及全基因组范围转录因子筛选获得了调控花青苷合成的两个新基因PyMADS18和PyMYB9，但是其功能及作用机制尚不明确。本项目拟通过对梨色泽突变体（红色/绿色）的PyMADS18和PyMYB9基因全长及启动子序列比较，明确序列结构变化对色泽发育的影响；并通过烟草和梨愈伤组织的RNAi及过量表达转基因鉴定相应的生物功能；同时，利用酵母双杂交及免疫共沉淀等技术分析转录因子互作蛋白和调控基因，从而明确克隆的两个新转录因子调控网络以及对花青苷合成结构基因的调控机制。本研究不仅有利于阐明梨花青苷合成调控的关键因子，完善果皮色泽发育和调控的科学理论，同时也将为推动红皮梨分子遗传改良提供极有价值的指导和必要的基因资源。

Red pear is currently attractive by customers and possessing extensive development potential in market. As we know,the red-skin color of pear fruit is caused by biosythesis and accumulation of anthocyanins. We founded that transcription factors (TFs) which regulate anthocyanin biosynthesis were important effectors for coloration of pear fruit. In the former research, we obtained the new TFs PyMADS18 and PyMYB9, which effectes anthocyanin biosynthesis significantly,by cDNA-AFLP and whole genome-wide TFs selection. However, the function and regulation mechanism of new genes are still unclear. In this project, the relationship between TF genes mutant and coloration of pear will be study through the comparation of whole length sequence of gene and promoters; the gene function will be identified by using over-expression and RNAi silencing in tobacoo plant and callus of pear;In addition, the proteins which interact with the PyMADS18 and PyMYB9 transcription factor and regulated genes will be screened by using the yeast two-hybrid system and co-immunoprecipitation techniques,then the regulatory network and regulation mechanism of new TF genes will be explored and declared. The results will not only help to elucidate the key factors which regulate the anthocyanin biosynthesis and complete the theory of fruit color development, but also help to promote the process of molecular breeding in red pear and provide necessary gene resources.