类钙调磷酸酶B亚基蛋白（CBLs）作为近年来新发现的一类植物Ca2+传感器蛋白，能与其互作的激酶CIPKs选择性特异结合，在调控植物响应非生物胁迫信号转导中发挥重要的作用。低温冷（冻）害是严重危害寒地葡萄栽培生产的主要逆境限制因子，有关葡萄CBL-CIPK参与低温胁迫调控的确切机制尚未阐明。本项目在前期对高抗寒山葡萄转录组测序获得CIPKs基因克隆、表达特性鉴定的基础上，拟挖掘参与低温胁迫的CBL-CIPK特异组合；明确其参与低温胁迫的功能特性，构建重要候选CIPK过量与RNAi表达载体，研究其在欧洲葡萄中过表达/被抑制与抗寒性的关系；结合cDNA-AFLP、CoIP与组学分析，初步构建特异CBL-CIPK调控的低温胁迫应答网络。课题的实施不仅有助于深层次理解山葡萄CBL/CIPK信号转导系统在抗寒代谢中的作用，而且对于通过分子育种解决制约我国宁夏葡萄产业存在的冷（冻）害问题具有现实意义。

Calcineurin B-like proteins (CBLs) represent a family of newly emerging plant-specific Ca2+ sensors, which could selectively interplay with their respective kinase effectors (CBL-interacting protein kinases, CIPKs), thus playing critical roles in response to signaling transduction of various abiotic stresses. Chilling/Freezing damage is a key environmental limitation factor that adversely affects grapevine cultivation and production in cold regions. The exact regulatory mechanism of the CBL-CIPK complexes in grapevine in response to cold stress remains to be elucidated. Based on the cloning and expression characteristics of CIPKs from our previous RNA-seq data, the specific CBL-CIPK signaling components that involved in cold stress will be identified and functional characterized in this project. We will further characterize the roles of the key candidate CIPKs in stable integrated transgenic V. vinifera plants by either overexpressing or inhibiting the target gene, thereby exploring their connection with cold hardiness. Combined with cDNA-AFLP, CoIP and omics analysis, a regulatory signaling network model of specific CBL-CIPK components in cold stress will be constructed in V. amurensis. The implementation of this project will not only help in understanding in-depth mechanistic interplay of CBL/CIPK mediated signaling pathway associated with cold tolerance in grapevine, but also possess a realistic meaning in resolving the existing chilling/freezing damage problem that limits the development of Ningxia grapevine industry in China.