As one of the key enzymes in the oxidative pentose phosphate pathway, glucose-6-phosphate dehydrogenase (G6PDH) plays a role in response to abiotic stresses and pathogenesis. Here, a full-length cDNA was obtained, designed as ScG6PDH from sugarcane. The ScG6PDH gene is 1,646 bp long with a 1,524-bp long ORF encoding 507 amino acid residues. Analysis of a phylogenetic tree indicated that this gene is a member of the cytosolic G6PDH gene family, which is consistent with results from a subcellular localization experiment. Based on a real-time quantitative RT-PCR performed under salt, drought, heavy metal (CdCl2) and low temperature (4 °C) treatments, the transcription levels of the ScG6PDH gene were higher compared with transcription levels where these treatments were not imposed, suggesting a positive response of this gene to these environmental stresses. Furthermore, G6PDH activity was stimulated under 4 °C, CdCl2, NaCl and PEG treatments, but the increments varied with treatment and sampling time, implying positive response to abiotic stresses, similar to the transcript of the G6PDH gene. Ion conductivity measurements and a histochemical assay provided indirect evidence of the involvement of the ScG6PDH gene in defense reactions to the above-mentioned abiotic stresses.