‘金坠梨’自交亲和性突变的分子机制研究

‘Jin Zhui’ is a spontaneous self-compatible mutant of ‘Ya Li’. Recent studies suggest that the pollen-part mutation of self compatible ‘Jin Zhui’ is associated with non S-locus PPM, which is unlinked to the S-locus. This project would identify the non S-locus PPM by using the method comprised proteomics and bioinformatics approaches. Firstly, we would identify non S-locus unknown gene in complete genome by bioinformatics approaches. And then we would design primers and clone the identified genes, according to the coding gene sequences. We would detect the expression pattern of genes between ‘Jinzhui’ and ‘YaLi’. Secondly, By using Two-dimensional fluorescence difference gel electrophoresis, we want to find the differentially expressed protein. We finally identify the non S-locus PPM by understanding the data from comprised proteomics and bioinformatics approaches. Thirdly, we would carry on the functional complement experiments, which would be done using pollen culture in vitro, to verify the function of non S-locus gene. In the end, we would discuss the correlation between the mutant non S-locus gene and the abnormal meiosis of pollen mother cells from ‘Jin Zhui’. The expected results woud provide not only new insight into the mechanism of self incompatibility of Rosaceae but also an important self compatible material for breeding.

‘金坠梨’是‘鸭梨’自交亲和性芽变品种。研究表明，‘金坠梨’自交亲和性突变与其花粉部分非S因子功能丧失密切相关。本项目拟通过利用基因组学和蛋白组学的方法，鉴定导致‘金坠梨’自交亲和性突变的非S因子。首先，利用梨基因组数据，通过生物信息学方法在全基因组中寻找潜在的非S基因，并设计引物，在‘金坠梨’和‘鸭梨’中克隆相关基因，通过RT-PCR检测克隆到的非S基因在‘金坠梨’和‘鸭梨’中表达模式是否相同；其次，利用双向荧光差异凝胶电泳技术，找到‘金坠梨’和‘鸭梨’花粉中有差异的蛋白位点进行质谱分析，通过整合基因组和蛋白组数据揭示导致‘金坠梨’自交亲和性突变的非S基因。同时，通过功能互补实验，利用体外花粉培养体系验证突变的非S基因功能。最后探讨，‘金坠梨’突变的非S基因与其花粉母细胞减数分裂异常之间的关系。预期结果将为蔷薇科果树自交不亲和机理研究提供新的理论依据，为果树亲和性育种提供重要材料。