苹果MdARF-MdHYL1-MdMYB88/MdMYB124-MdSNF2调控网络解析及抗寒功能鉴定

Low temperature affects apple quality and productivity in Loess Plateau, hence identification of freezing important genes is required for breeding of freezing-tolerant apple by gene engineering. Our previous research showed that MdMYB88/MdMYB124 is a positive regulator for apple freezing tolerance, which also positively regulates MdSNF2 expression. In addition, MdMYB88/MdMYB124 interacts with MdHYL1, a positive regulator for apple freezing tolerance, which was directly regulated by MdARF. Based on these results, we are going to confirm the binding of MdARF to promoter of MdHYL1 by EMSA and ChIP-qPCR; to identify biological function of MdHYL1,MdARF,MdSNF2 under freezing stress; and to examine the molecular mechanisms of the interaction between MdMYB88/MdMYB124 and MdHYL1.The goal of this project is to characterize the function and molecular mechanisms of MdARF-MdHYL1-MdMYB88/MdMYB124-MdSNF2 in apple in response to freezing stress. Our results will provide theoretic foundation for apple in response to low temperature, and new genes for breeding of apple with freezing tolerance by gene engineering.

低温影响黄土高原产区苹果质量与产量，耐寒基因的发掘是通过基因工程创制耐寒苹果种质的重要前提。课题组前期发现苹果MYB转录因子MdMYB88/MdMYB124具有抗寒功能、正调控MdSNF2，并与MdHYL1互作。前期研究还发现了MdHYL1的上游调控因子MdARF。在此基础上，本项目拟进一步采用EMSA及ChIP-qPCR验证MdARF对MdHYL1启动子的结合；拟通过创制转基因苹果植株确定MdHYL1、MdARF、及MdSNF2在苹果抗寒中的生物学功能；并明确MdMYB88/MdMYB124与MdHYL1互作的机制。本项目的主旨是解析苹果MdARF-MdHYL1-MdMYB88/MdMYB124-MdSNF2调控网络及其抗寒功能，为阐明苹果低温应答的调控网络奠定理论基础，并为开展苹果抗寒基因工程和创制苹果新种质提供重要的基因资源。

低温影响黄土高原产区苹果质量与产量，耐寒基因的发掘是通过基因工程创制耐寒苹果种质的重要前提。课题组前期发现苹果MYB转录因MdMYB88/MdMYB124具有抗寒功能、正调控MdSNF2，并与MdHYL1互作。前期研究还发现了MdHYL1的上游调控因子MdARF。在此基础上，本项目进一步采用EMSA及ChIP-qPCR验证了MdARF对MdHYL1启动子的结合；通过创制转基因苹果植株确定了MdHYL1、MdARF、及MdSNF2在苹果抗寒中的生物学功能；并明确了MdMYB88/MdMYB124与MdHYL1互作的机制。项目解析了苹果MdARF-MdHYL1-MdMYB88/MdMYB124-MdSNF2调控网络及其抗寒功能，阐明苹果低温应答的调控网络奠定了理论基础，并为开展苹果抗寒基因工程和创制苹果新种质提供了重要的基因资源。