The effect of exogenous nitric oxide (NO) on chilling injury to banana fruit was investigated. Banana fruit was treated with NO donor sodium nitroprusside of 0.05 mM at 20 °C for 10 min and then stored at 7 °C for up to 20 days. Banana fruit treated with NO sustained a lower chilling injury index and higher firmness and kept lower electrolyte leakage and malondialdehyde content than the control. Further investigation showed that NO treatment enhanced activities of guaiacol peroxidase, ascorbate peroxidase, and glutathione reductase compared to the control. It also maintained higher ascorbic acid, reduced glutathione content, and total antioxidant capacity but reduced hydrogen peroxide and superoxide anion to lower levels compared to control fruit during storage. NO treatment significantly enhanced the accumulation of total phenolics and proline, which resulted from the increased activities of phenylalanine ammonia-lyase and Δ¹-pyrroline-5-carboxylate synthetase and decreased proline dehydrogenase activity. We proposed that the enhanced chilling tolerance induced by NO treatment may result from the reduction of oxidative stress and proline accumulation.