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β-aminobutyric acid (BABA) is a new environmentally friendly agent to induce disease resistance by priming of defense in plants. However, molecular mechanisms underlying BABA-induced priming defense are not fully understood. Here, comprehensive analysis of priming mechanism of BABA-induced resistance was investigated based on mango-Colletotrichum gloeosporioides interaction system using iTRAQ-based proteome approach. Results showed that BABA treatments effectively inhibited the expansion of anthracnose caused by C. gleosporioides in mango fruit. Proteomic results revealed that stronger response to pathogen in BABA-primed mango fruit after C. gleosporioides inoculation might be attributed to differentially accumulated proteins involved in secondary metabolism, defense signaling and response, transcriptional regulation, protein post-translational modification, etc. Additionally, we testified the involvement of non-specific lipid-transfer protein (nsLTP) in the priming acquisition at early priming stage and memory in BABA-primed mango fruit. Meanwhile, spring effect was found in the primed mango fruit, indicated by inhibition of defense-related proteins at priming phase but stronger activation of defense response when exposure to pathogen compared with non-primed fruit. As an energy-saving strategy, BABA-induced priming might also alter sugar metabolism to provide more backbone for secondary metabolites biosynthesis. In sum, this study provided new clues to elucidate the mechanism of BABA-induced priming defense in harvested fruit.