低温影响植物的生长发育、产量和生态分布，而对于热带起源的香蕉（包括香牙蕉、大蕉等）来说更是如此。近年来频繁发生的倒春寒已给我国香蕉产业造成了巨大的经济损失。生产实践中发现大蕉的耐冷性显著强于主栽品种香牙蕉，申请人前期的研究表明MAPK级联-ICE1途径，特别是MKK2、MAPK3和ICE1在大蕉耐冷过程中起着关键的作用，然而目前对其互作调控机理尚不清楚。本项目拟以超表达mCherry-MKK2/MAPK3/ICE1转基因大蕉幼苗为材料，采用最新的交联质谱技术高通量和高灵敏度地鉴定低温处理下与大蕉MKK2、MAPK3和ICE1直接和间接互作的蛋白，然后利用酵母双杂和双分子荧光互补实验进行验证，最后通过在香蕉中超表达和突变重要互作蛋白编码基因解析其生物学功能。本项目研究结果不但可以深入揭示MKK2、MAPK3和ICE1在大蕉抗寒过程中的作用机理，也可为香蕉抗寒分子育种提供有效的基因资源。

Low temperature affects the growth, development, productivity and geographical distribution of plants, especially for bananas (included Cavendish, Dajiao and so on) originated in the tropics. Although they are mainly cultivated in tropical and subtropical regions, cold spells often occur during winter or early spring, which can have serious impact on fruit quality and yield, and also cause in some incidents the destruction of banana plantations. The cold resistance of Dajiao was significantly higher than that of main varieties (Cavendish), and our previous studies showed that the MAPK cascade -ICE1 pathway, especially MKK2, MAPK3 and ICE1, played a critical role in Dajiao cold resistance, however, their interaction regulation mechanism is unclear. This research will take transgenic Dajiao seedlings of overexpressing mCherry-MKK2, mCherry-MAPK3 and mCherry-ICE1 as materials, to identify the direct and indirect interaction proteins of MKK2, MAPK3 and ICE1 under low temperature treatment by the latest high throughput and high sensitivity cross-linking mass spectrometry technology, then the Yeast Two-Hybrid system and Bimolecular Fluorescence Complementation (BiFC) used to verify their interaction, and its biological function will be disclosed by creating overexpress plants or mutated-plants using CRISPR/Cas9 system. The outcome of the proposed research will be helpful for in-depth understanding the molecular networks of cold-resistant process that modulated by MKK2, MAPK3 and ICE1 in Dajiao, but also provide key candidate genes for cold-resistant banana breeding.