果刺颜色是黄瓜非常重要的外观品质性状。前人研究表明CsMYB60为黑刺基因B的最佳候选基因，但其生物学功能尚未确证，其调控黑刺形成的分子机理亦不清楚。本课题组前期工作明确了黑刺着色物质主要为类黄酮，发现其在黄瓜根腐病抗性中起重要作用，并证实了CsMYB60是调控黑刺颜色形成的关键因子。本项目拟利用酵母单杂交、GUS瞬时表达系统，明确该基因是否直接调控类黄酮合成途径中结构基因的表达；利用酵母双杂交筛选CsMYB60互作蛋白，用 BiFC及CoIP确证后，在果刺瞬时表达系统中验证其生物学功能，然后明确CsMYB60及其互作蛋白对类黄酮合成结构基因Cs4CL的调控模式；将CsMYB60和Cs4CL进行遗传转化，对转基因植株进行分子鉴定、表型分析、类黄酮测定以及转录组分析，进而解析CsMYB60调控黄瓜黑刺类黄酮积累的分子机理，为黄瓜品质及抗病分子育种提供理论依据。

The spine color of cucumber is one of the important quality traits. Previous study showed that CsMYB60 was the best candidate gene for the B locus conditioning the pigmentation of black spines, but its biological function has not yet been validated and the mechanism of the coloration of black spines regulated by CsMYB60 still remains unclear. Our previous results indicated that the pigments in black spines are mainly flavonoids which play vital roles in resistance against cucumber root rot, and proved that CsMYB60 is a key factor for the pigmentation of black spines. In this project, we will clarify whether CsMYB60 directly regulates structure genes required for flavonoid biosynthesis by yeast one-hybrid and GUS transient expression system. The interacting proteins of CsMYB60 will be screened by yeast two-hybrid system, and verified with BiFC and CoIP methods, and then we will explore their biological functions with transient expression system in fruit spines. To verify the function of CsMYB60 and Cs4CL, these two genes will be transformed into the cucumber with white spine, then molecular identification, phenotypic analysis, component and quantitative analysis of flavonoids, transcriptomic analysis will be done in the transgenic plants. So, this project will elucidate the molecular mechanism of flavonoid biosynthesis controlled by CsMYB60 in cucumber black spines, which will provide theoretical basis for molecular breeding of quality and disease resistance in cucumber.