Abstract: Banana is one of the very important economic crops in tropical area of China, and the banana industry has been seriously affected by chilling often. Clarifying the molecular mechanism of banana plants in response to low temperature stress could provide the research basis for cultivating and screening relatively cold resistant banana varieties. Comparative proteomics was applied to analyze the differential expression proteins of banana seedling leaves which were exposed at 7℃ for 0, 3, 5, 7 days, and the expression of 85 protein spots was found to be obviously affected by low temperature, finally 82 proteins out of them were identified by mass spectrum. These identified proteins were mainly involved in protein synthesis process, protein posttranslational modification, carbohydrate and energy metabolism, and photosynthesis. The results suggested that the banana seedling leaf might regulate carbon and energy metabolism through the expression pattern change of photosynthesis which was related to enzymes, and carbohydrate which was associated with energy metabolism enzyme to ensure the supply and absorption of carbon nutrients, and also improve the ability of low temperature resistance by superoxide dismutase scavenging system. The results could provide new clues for the study of the molecular mechanism of low temperature resistance of banana, and new candidate proteins for the breeding of cold resistant strain of banana.