红梨着色模式多样，现有红梨着色机制的研究多集中在花青苷合成相关基因克隆，表达分析和功能鉴定的传统思路上，对这些基因转录调控机制研究极为薄弱，所涉及的研究材料也相对单一，难以从全局上认识不同着色类型红梨着色的分子调控机制。本研究从启动子序列变异和DNA甲基化分析两个角度系统研究红梨花青苷合成相关基因转录调控机制，拟以包括东方梨、西方梨和特殊类型等具有不同着色模式的典型红与非红梨为试材，分离其花青苷合成相关基因编码区和启动子，并对这些基因序列变异与色泽变异进行关联分析，找出与色泽变异相关的序列变异，再利用双荧光素酶检测、启动子删截、酵母双杂交、凝胶阻滞分析和转基因等实验手段对该序列进行功能鉴定分析，对于无法从基因序列变异角度解释的色泽变异现象，则对其进行DNA甲基化水平检测。研究成果有助于阐明红梨花青苷合成调控机制，同时可进一步丰富果实花青苷生物合成的分子机制的理论。

Red pears belonging to different species with different genetic backgrounds and coloration patterns may have the distinct transcriptional regulation for related anthocyanin biosynthesis genes compared with apples and grape. However, more attention was payed on the isolation, expression and function analysis of related genes in the study on molecular mechanism of coloration in red pears, few have carried out on the mechanism of transcriptional regulation. What's more, the limited and single material in the previous studies related to red pears would not be favorable to the comprehensive understanding of molecular regulation mechanism of different kinds of red pears. Therefore, in this project, typical red pears including Oriental pear, Occidental pear and some special types with different coloration patterns will be used to study transcriptional regulation for related anthocyanin biosynthesis genes. The promoter region will be isolated for analyzing the correlation between the sequence variations in the promoter region and peel phenotypes of red pears. The dual luciferase assay, promoter deletion analysis, electrophoretic mobility shift assay, yeast two-hybrid assay, transgenic technology will be further used for the analysis of their function. Methylation levels of these genes will also be investigated to understand molecular mechanism for coloration unrelated with the sequence variations in the promoter region. The research will be of help to clarify the regulation mechanism of anthocyanin biosynthesis in red pears, and also to enrich the theory of molecular mechanism of fruit anthocyanin biosynthesis.