前期工作基于红花转录组及代谢组数据库，以不含羟基红花黄色素A(hydroxysafflor yellow A，HSYA)的红花W品系为对照，通过对高HSYA的Y品系花冠时序性差异表达基因与时序性累积黄酮类代谢物的关联分析，获得了可能参与HSYA生物合成的功能基因。进而以黄酮类代谢物为配体，以基因预测的蛋白酶为受体，采用反向分子柔性对接策略，预测出了HSYA生物合成途径及其关键基因：1个CYP450、3个Reductases和2个 UGTs。本研究拟在分析候选基因时空表达及HSYA时空分布特征的基础上，通过构建Crispr/cas9基因敲除载体，以花粉管通道法遗传转化红花Y品系，通过对基因敲除突变体从基因表达到代谢物的分析，验证其生物学功能；通过构建异源表达载体，鉴定酶的催化活性。最终明确HSYA生物合成关键基因的功能，解析其代谢途径。为后续HSYA的合成生物学研究提供重要科学依据。

In the early stage, basing on safflower transcriptome and metabolomics database, and as negative control of HSYA-absent W line, we successfully performed the correlated analysis between differential genes and flavonoids metabolites changes depending on the timeline of florets developmental stages, and obtained potential functional genes participated in HSYA biosynthesis. Furtherly, by adopting of Reverse Flexible Docking Strategy (RFDS), the flavonoid metabolites used as ligands and putative protein enzymes as receptors, it’s predicted that the biosynthetic pathway of HSYA and key genes: 1 CtCYP450, 3 reductases as well as 2 CtUGTs. To valid their biological functions, at first, through constructing Crispr/cas9 plasmid and transforming Y line by the method of pollen-tube, we are attempting to figure out gene expression and metabolites accumulation changes. On the other hand, heterologous protein expressions would be carried out and enzymatic activities be identified in vitro. In the end, we would conform key genes’ functions and specify HSYA biosynthetic pathway. This study would provide significant foundation for following HSYA synthetic biology studies.