苹果高类黄酮优异种质创制及其类黄酮代谢机理的研究

创制高类黄酮优异种质、培育易着色的高类黄酮苹果等特色多样化品种是苹果产业的重要技术需求。本申请在高类黄酮优异种质CSR6R6创制及其杂种后代分离群体构建以及通过RNA-seq与基因表达分析获得HsfA8和TMT1/2等候选基因的基础上，进一步通过初选、复选和决选程序，选育高类黄酮苹果新品种；以CSR6R6及其高类黄酮苹果新品系为试材，通过基因克隆和生物信息学分析等研究候选基因结构特征和时空表达特性及其与参试品种果实类黄酮含量变化的关系；利用酵母双杂交及EMSA等技术研究候选基因与FLS等相关基因在蛋白-蛋白和蛋白-DNA水平上的互作特点及调控作用，并进行转基因功能验证，研究热激转录因子、糖转运蛋白和激素信号因子与高类黄酮苹果类黄酮代谢的关系和作用机理，从而探讨高类黄酮苹果优异种质的类黄酮代谢机理，为丰富苹果资源及苹果类黄酮代谢机理的理论体系提供基本资料，并为高类黄酮苹果育种提供科学依据。

The creation of excellent apple germplasm with high flavonoid and easy coloring and the breeding of characteristic varieties of high-flavonoid apples are important technical need for apple industry. This application is based on the creation of high flavonoid germplasm ‘CSR6R6’, the construction of hybrid population and the obtainment of candidate genes such as HsfA8, TMT1/2 and ARF3 by phenotypic analysis, RNA-seq and gene expression analysis. Further through the primary selection, the second selection and the final selection procedures, new functional apple cultivars were bred. The germplasm of CSR6R6 and its hybrid progeny lines were took as materials. We studied the structural characteristics and temporal and spatial expression of candidate genes and their relationship with the content of flavonoids in fruits; Yeast two hybrids, EMSA and other techniques will be used to study the protein-protein and protein-DNA interaction characters and regulation functions between candidate genes and structure genes, and transgenic function verification will be performed. The relationship between heat shock transcription factor, glucose transporter, auxin signaling factor and flavonoid metabolism was studied in high-flavonoid apples to investigate their flavonoid metabolism mechanism, so as to provide basic data for the enriching apple resources and the theoretical system of flavonoids metabolism and provide the scientific basis for the breeding of high-flavonoid apples.