花香是观赏植物的重要品质之一，最近的研究表明PhABCG1转运蛋白在调控矮牵牛花香物质释放过程中发挥着重要作用，但其转录调控机制目前尚不清楚。申请者前期通过酵母单杂交筛选获得了与转运蛋白基因PhABCG1启动子结合的锌指蛋白PhZPT2-5，但其具体调控机制未知。本项目拟利用双荧光素酶系统和EMSA技术验证PhZPT2-5蛋白与PhABCG1启动子互作情况。在此基础上，分析PhZPT2-5基因的时空表达规律，并利用过表达和RNAi等手段探究PhZPT2-5基因对不同花香成分释放的影响。最后利用TurboID技术筛选与PhZPT2-5蛋白互作的转录因子，并利用酵母双杂交和荧光素酶互补系统进行验证。该研究不仅可以揭示PhZPT2-5通过调控PhABCG1基因的转录进而影响矮牵牛花香物质释放的分子机制，还可为从分子层面改良观赏植物花香品质奠定重要的理论基础。

Floral scent is one of the important characteristics in ornamental plants. Previous research has revealed that PhABCG1 transporter protein play essential roles in the emission process of floral volatile organic compounds in petunia. However, the mechanism of transcription regulation is not clear at present. In our previous study, a C2H2-type zinc finger protein PhZPT2-5 binding to the promoter of PhABCG1 was obtained by yeast one hybrid screening, but its specific regulation mechanism is unknown. In this project, dual luciferase reporter system and EMSA technology will be used to verify the interaction between PhZPT2-5 protein and the promoter of PhABCG1. What’s more, the temporal and spatial expression patterns of PhZPT2-5 gene will be analyzed, and the effect of PhZPT2-5 gene on the emission of different floral volatile organic compounds will be explored in PhZPT2-5 overexpressing and RNAi transgenic plants. Finally, the transcription factors interacting with PhZPT2-5 protein will be screened by TurboID technology, and will be verified by yeast two hybrid and luciferase complementary imaging assay. This study can not only reveal the molecular mechanism of the C2H2-type zinc finger protein PhZPT2-5 affecting the emission of floral volatile organic compounds by regulating PhABCG1 gene transcription in petunia, but can also lay an important theoretical foundation for improving the floral scent quality in ornamental plants at the molecular level.