Low temperature is one of the most prominent environmental factors affecting plant growth. As a deciduous arboreal tree, Acer palmatum has considerable ornamental and economic value; however, the molecular mechanisms underlying cold stress regulation in this species have yet to be determined. In this study, we performed Illumina high-throughput sequencing of 18 libraries obtained from A. palmatum subjected to cold treatment and subsequently identified a differentially expressed R2R3-MYB family gene, ApMYB77 , which was then cloned and functionally characterized. The expression of ApMYB77 was induced by cold and drought treatments, and overexpression in A. thaliana enhanced the freezing tolerance (− 9 °C for 6 h) of transgenic plants compared with that of wild-type plants. The survival rates of transgenic plants (89% and 92%) were significantly higher than the wild-type plants (52%). Moreover, the transcript abundances of CBF-dependent regulatory pathway genes ( AtCBF1 , AtCBF2 , AtCBF3 , AtCBF4 , AtCOR6.6A , AtCOR15B , AtCOR78 , AtCOR414 , and AtKIN1 ) were found to be significantly up-regulated in the transgenic lines. Enhanced abscisic acid (ABA)-dependent drought tolerance in transgenic plants was a further consequence of the overexpression of ApMYB77 following treatment of 15% polyethylene glycol for 4 d, compared with that in wild-type plants. In contrast, the expressions of genes ( AtANAC072 , AtDREB2A , AtERD1 , AtMYB2 , AtRD20 , and AtRD29A ) positively regulated by ABA were activated. Overall, the findings of this study indicate that ApMYB77 confers both freezing and drought tolerances.