In order to explore the role of superoxide dismutase (SOD) gene in maize stress resistance, we selected two maize seedlings with distinct differences in drought resistance as experimental materials. The sequence structure and expression patterns of ZmSOD gene under drought stress were analyzed by RT-PCR, quantitative real-time PCR (qRT-PCR) and two-dimensional gel electrophoresis (2-DE) coupled with matrix assisted laser desorption ionization/time of flight mass spectrometry (MALDI-TOF). The results showed that: (1) The full-length cDNA sequences of SOD gene were cloned successfully from the leaves of drought-sensitive Denghai 605 (DH605) and drought-resistant Liyu 35 (LY35), respectively. The opening reading frame (ORF) of ZmSOD1 gene in DH605 was 456 bp in length, encoding 151 amino acids. The isoelectric point of ZmSOD1 protein was 5.76, and its molecular weight was 15.05 kD. The ORF of ZmSOD2 gene in LY35 was 459 bp in length, encoding 152 amino acids. The isoelectric point of ZmSOD2 protein was 5.65, and its molecular weight was 15.11 kD. ZmSOD1 and ZmSOD2 are hydrophilic stable proteins, both containing Cu/Zn-SOD domain without signal peptide and transmembrane domain at the N-terminal. Homology and phylogenetic analysis showed that ZmSOD was highly homologous to FmCu/Zn-SOD2, and the similarity was as high as 96.05%. (2) Under drought conditions, the transcription level of ZmSOD1 in DH605 was significantly decreased, and that of ZmSOD2 in LY35 was significantly increased. The 2-DE coupled mass spectrometry showed that the expression level of ZmSOD1 in DH605 was significantly decreased, the expression level of ZmSOD2 in LY35 was not significantly changed, but the expression level of Mn-SOD was significantly increased. (3) Correlation analysis showed that the transcription level of ZmSOD1 in DH605 leaves was marked positive correlated with its protein abundance and SOD activity, and the transcription level of ZmSOD2 in LY35 leaves was positively correlated with SOD activity. The results of this study provided a basis for further exploring the mechanism of SOD gene in regulating maize resistance and stress response.