AtNDPK2 is involved in transcriptional regulation in response to pathogen and abiotic stresses. AtNDPK2-expressing transgenic rice plants showed regulation of the marker genes for chilling and oxidative stresses. In the present study, we produced AtNDPK2-overexpressing transgenic rice lines using the co-transformation method. Morphologically, the transgenic plants, compared with the control plants, were growth retarded. We investigated how AtNDPK2 overexpression influences the response of rice plants to marker genes related to chilling and ROS stress. The accumulation of transcripts of pBC442 and pBC601, related to chilling stress, was induced in AtNDPK2-overexpressed rice plants. On further investigation, we found that OsAPX1-, OsAPX2-, and OsSodB-scavenging free-oxygen radicals, such as superoxide (O2-) and hydrogen peroxide (H(2)O(2)), could be induced in AtNDPK2-overexpressed rice plants. In particular, transcripts encoding pathogenesis-related (PR) proteins OsPR2 and OsPR4, as well as oxidative stress response proteins, were confirmed to change the gene expression in the transgenic rice plants. Together, these results suggest that AtNDPK2 plays a regulatory role in chilling and antioxidant signaling in plants.