以果实为产品器官的农作物开花早晚影响着产品收获期，弄清决定其开花早晚的因子并研究其调控机制具有重要意义。申请人前期研究发现，转录因子FaARF4具有调控草莓开花早晚的功能，因此本项目拟采用转录组测序和qRT-PCR技术，筛选FaARF4调控的下游靶基因，并借助酵母单杂交和EMSA试验，确定FaARF4与靶基因间的互作关系；通过FaARF4靶基因过量表达及沉默来分析其对草莓开花早晚的影响；利用FaARF4靶基因::GFP融合基因通过转化洋葱表皮细胞，进行ARF4靶蛋白的亚细胞定位；结合GUS组织染色分析转proARF4::GUS基因植株在不同激素种类、浓度等处理下启动子的转录活性，进而明确影响ARF4基因表达的因素。通过上述研究揭示FaARF4调控草莓开花的分子机理，为通过调控ARF4基因表达来提早或延晚草莓开花奠定基础，并可为其他果树作物开花机理研究提供理论参考。

Flowering time of crops with fruit as the product will affect harvest period. It is of great significance to find the factors that determine the flowering time and to identify their regulatory mechanism. Our previous studies have found that FaARF4 transcription factor can regulate the flowering time of strawberry. The present research program will find out the target genes regulated by FaARF4 with transcriptome sequencing and qRT-PCR. The interaction between FaARF4 and its target genes will be confirmed by yeast one-hybrid system and electrophoretic mobility shift assay (EMSA). Its function in the flowering time regulation will be analyzed by adopting over-expression and silence of ARF4 target genes in strawberries, respectively. The subcellular localization of FaARF4 target protein will be investigated by transforming target gene::GFP into onion epidermal cells. According to GUS staining analysis, the promoter activity of ARF4 gene will be elucidated in the transgenic plants with proARF4::GUS which will be treated with different types and concentrations of hormones, etc. And the factors affecting the expression of ARF4 gene will be further confirmed. The aim of this project is to reveal the molecular mechanism of FaARF4 gene in regulating strawberry flowering. The findings will lay a foundation for flowering promotion or inhibition of strawberry by regulating the expression of ARF4 gene. The results will also provide theoretical reference for the research on flowering mechanism of other fruit trees.