Cold stress is one of the most serious abiotic stresses that affects the growth and yield in rice. However, the molecular mechanism by which abscisic acid (ABA) regulates plant cold stress tolerance is not yet clear. In this study, we identified a member of the OsNCED (9-cis-epoxycarotenoid dioxygenase) gene family, OsNCED5, which confers cold stress tolerance in rice. OsNCED5 encodes a chloroplast-localized ABA biosynthetic enzyme and its expression is strongly induced by cold stress. Disruption of OsNCED5 by CRISPR/Cas9-mediated mutagenesis led to a significant decrease in ABA content and exhibited significant reduced cold stress tolerance at the seedling stage. Exogenous ABA restored the cold stress tolerance of the osnced5 mutants. Overexpression of OsNCED5 gene significantly improved the cold stress tolerance of rice seedlings. Moreover, OsNCED5 mainly regulates cold stress tolerance through regulating reactive oxygen species (ROS) homeostasis. Taken together, we identified a new OsNCED regulator involved in cold stress tolerance, and provided a potential target gene for enhancing cold stress tolerance in rice.