Heat stress is a major restrictive factor that suppresses rice production. In this study, we investigated the potential priming effect of exogenous abscisic acid (ABA) on heat tolerance in rice seedlings. Seedlings were pretreated with 10 μM ABA by root drenching for 24 h and then subjected to heat stress conditions of 40 °C day/35 °C night. ABA pretreatment significantly decreased leaf withering by 2.5– 28.5% and chlorophyll loss by 12.8–35.1% induced by heat stress in rice seedlings. ABA application also mitigated cell injury, as shown by lower malondialdehyde (MDA) content, membrane injury and expression of cell death-related genes OsKOD1 , OsCP1 and OsNAC4 , while expression of OsBI1 , a cell death-suppressor gene, was upregulated by ABA pretreatment. Moreover, ABA pretreatment improved antioxidant defense capacity, as shown by an obvious upregulation of ROS-scavenging genes and a decrease in ROS content (O 2 – and H 2 O 2 ) and downregulation of the OsRbohs gene. The application of fluridone, an ABA biosynthesis inhibitor, increased membrane injury and the accumulation of ROS under heat stress. Exogenous potent antioxidants (proanthocyanidins, PC) significantly alleviated leaf withering by decreasing ROS overaccumulation and membrane injury induced by heat stress. In addition, ABA pretreatment significantly superinduced the expression of ABA-responsive genes SalT and OsWsi18 , the ABA biosynthesis genes OsNCED3 and OsNCED4 , and the heat shock-related genes OsHSP23.7 , OsHSP17.7 , OsHSF7 and OsHsfA2a . Taken together, these results suggest that exogenous ABA has a potential priming effect for enhancing heat stress tolerance of rice seedlings mainly by improving antioxidant defense capacity and heat shock-related genes.