Litchi (Litchi chinensis) is an economically important fruit tree in southern China and is widely cultivated in subtropical regions. However, irregular flowering attributed to inadequate floral induction leads to a seriously fluctuating bearing. Litchi floral initiation is largely determined by cold temperatures, whereas the underlying molecular mechanisms have yet to be identified. In this study, we identified fourCRT/DRE BINDING FACTORS(CBF) homologs in litchi, of whichLcCBF1,LcCBF2andLcCBF3showed a decrease in response to the floral inductive cold. A similar expression pattern was observed for theMOTHER OF FT AND TFL1homolog (LcMFT) in litchi. Furthermore, both LcCBF2 and LcCBF3 were found to bind to the promoter ofLcMFTto activate its expression, as indicated by the analysis of yeast-one-hybrid (Y1H), electrophoretic mobility shift assays (EMSA), and dual luciferase complementation assays. Ectopic overexpression ofLcCBF2andLcCBF3inArabidopsiscaused delayed flowering and increased freezing and drought tolerance, whereas overexpression ofLcMFTinArabidopsishad no significant effect on flowering time. Taken together, we identified LcCBF2 and LcCBF3 as upstream activators ofLcMFTand proposed the contribution of the cold-responsive CBF to the fine-tuning of flowering time.