Plant mitogen-activated protein kinase (MAPK) cascades play a pivotal role in a range of biotic and abiotic stress responses. In this study, we isolated a novel group D MAPK gene, ZmMPK17, from maize (Zea mays L.). ZmMPK17 is localized mainly to the nucleus and its C-terminal domain extension is believed to be essential for this. Northern-blot analysis indicated that ZmMPK17 transcription is involved in response to exogenous signaling molecules such as abscisic acid, hydrogen peroxide, salicylic acid, jasmonic acid and ethylene and induced by low temperature and osmotic stress. Hydrogen peroxide and Ca²⁺ mediate PEG-induced downregulation of ZmMPK17 at transcription level and Ca²⁺ also mediates low temperature-induced expression of ZmMPK17. Overexpression of ZmMPK17 in tobacco (Nicotonia tobaccum) accumulated less reactive oxygen species under osmotic stress by affecting antioxidant defense systems. Transgenic tobacco exhibited enhanced tolerance to cold by means of an increased germination rate, and increased proline and soluble sugar levels relative to control plants. The transcription levels of NtERD10 genes were higher in ZmMPK17-overexpressing lines than in control plants under cold and osmotic stress conditions. ZmMPK17-overexpressing plants displayed enhanced resistance to viral pathogens, and the expression of the pathogenesis-related gene PR1a was significantly increased, indicating that ZmMPK17 might be involved in SA-mediated pathogen defense-signaling pathways.