低温是限制柑橘产业发展的主要因素之一，提高柑橘抗寒性是育种的重要目标。我们前期研究从枳低温表达谱中鉴定出一个快速且强烈响应低温的串联C3H型锌指蛋白（PtrTZF1），并验证了其抗寒功能。但它提高抗寒性的分子机制和涉及的调控网络不明晰。本项目将构建超表达和基因编辑载体转化枳，进一步明确PtrTZF1的抗寒功能；在此基础上，对超表达植株进行RNA-Seq、RIP-Seq和ChIP-Seq分析，在转录和转录后水平上全面筛选PtrTZF1调控的靶基因，验证后获得PtrTZF1的调控元（regulon）；此外，构建枳低温cDNA文库，采用酵母双杂交筛选分离PtrTZF1的互作蛋白，并分析其生物学功能。本项目是已有研究的深入和延续，将为柑橘抗寒育种提供重要的基因资源，同时有利于揭示PtrTZF1的作用机制，为解析以它为节点的低温胁迫应答网络提供理论依据，因而具有重要的理论意义和潜在应用价值。

Low temperature is one of the main factors limiting the development of citrus industry, and improving the cold resistance is one crucial objective of citrus breeding. In the previous study, a tandem CCCH-type zinc-finger protein (PtrTZF1) was identified from the low-temperature treated expressional profile, and demonstrated functioning in cold resistance. However, the molecular mechanisms and regulatory network involved in its improvement of cold resistance remain elusive. In this project, an overexpression and a gene editing vector will be constructed and transformed into trifoliate orange to further clarify the cold resistance function of PtrTZF1. Furthermore, RNA-Seq, RIP-Seq and ChIP-Seq will be applied in overexpression plants to screen the target genes regulated by PtrTZF1 comprehensively at the transcriptional and post-transcriptional level, and the regulon of PtrTZF1 will be obtained after verification. In addition, a low-temperature cDNA library of trifoliate orange will be constructed, and the proteins interacting with PtrTZF1 will be screened and isolated by yeast two-hybrid. Then the biological function of the interacted proteins will be analyzed. This project is an in-depth and continuous work based on the previous research, which will provide valuable gene resources for cold resistant breeding of citrus, and help to reveal the mechanism of PtrTZF1, and provide theoretical basis for the investigation of the response network of cold stress with it as the node. Therefore, it takes important theoretical significance and potential application value.