香蕉果实后熟过程中，其品质、风味逐渐形成，为人类健康提供了必要的维生素、氨基酸、矿质营养等。申请人研究发现生长素对香蕉的后熟进程有显著的正调控作用。ARF作为生长素信号中重要的转录因子，可能是生长素信号与乙烯信号相互作用的桥梁。然而，其调控机制目前尚不清楚，靶基因还需进一步实验鉴定。本项目中拟以香蕉果实为材料，利用表达谱、蛋白组、Western blot、定量PCR等手段筛选出香蕉后熟进程中起关键作用的ARF，并构建植物超表达载体，利用农杆菌介导的转化体系转化番茄，验证ARF的基因功能。随后纯化ARF的单克隆抗体，通过多种蛋白互作技术分析与ARF相互作用的蛋白质，同时利用ChIP-Seq和酵母单杂交等手段鉴定ARF调控的下游靶基因。最后依据现有表达谱数据和已报道的文献数据构建生长素信号和乙烯信号的调控路径图，解析ARF在香蕉后熟过程中的调控机制，同时完善生长素信号和乙烯信号相互作用的模式。

Banana fruit ripening is a complex, genetically programmed process involving remarkable changes in the color, texture, aroma and flavor of the fruit, which provides the necessary vitamins, amino acids, minerals and other nutrients for human health. Our previous studies revealed that auxin response factor (ARF) might be not only a key transcription factor in auxin signal but also an important link in auxin signal with ethylene signal, which actively involve in the regulation of banana fruit ripening. However, the molecular mechanism is largely unknown, and the downstream target genes of ARF need to be identified. In this project, key ARF associated with banana ripening will be screened using a variety of analysis, such as gene expression profiling, proteomics analysis, western blot and real-time quantitative PCR. Plant expression vector will be constructed, and transformed into tomato using agrobacterium-mediated transformation system. The gene function of ARF will be verified in tomato ripening process. Monoclonal antibody of ARF will be purified and proteins interacted with ARF will be analyzed through a variety of protein interactions technical analysis. Meanwhile, the downstream target genes of ARF will be identified by using ChIP-Seq and yeast one-hybrid technologies. Finally, the pathways of auxin signaling and ethylene signaling will be constructed based on the existing gene expression profiling data and reported data in other references. The crosstalk of these two signaling pathways will be analyzed, which might reveal the regulatory mechanism of ARF in bananas ripening, and improve the model of interaction between auxin and ethylene signaling.