Chilling stress seriously limits grain yield and quality worldwide. However, the genes and the underlying mechanisms that respond to chilling stress remain elusive. This study identified ABF1, a cold-induced transcription factor of the bZIP family. Disruption ofABF1impaired chilling tolerance with increased ion leakage and reduced proline contents, whileABF1over-expression lines exhibited the opposite tendency, suggesting that ABF1 positively regulated chilling tolerance in rice. Moreover, SnRK2 protein kinase SAPK10 could phosphorylate ABF1, and strengthen the DNA-binding ability of ABF1 to the G-boxcis-element of the promoter ofTPS2, a positive regulator of trehalose biosynthesis, consequently elevating theTPS2transcription and the endogenous trehalose contents. Meanwhile, applying exogenous trehalose enhanced the chilling tolerance ofabf1mutant lines. In summary, this study provides a novel pathway ‘SAPK10-ABF1-TPS2’ involved in rice chilling tolerance through regulating trehalose homeostasis.