The production of the marine trace gas dimethyl sulfide (DMS) provides 90% of the marine biogenic sulfur in the atmosphere where it affects cloud formation and climate. The effects of increasing anthropogenic CO2 and the resulting warming and ocean acidification on trace gas production in the oceans are poorly understood. Here we report the first measurements of DMS-production and data on growth, DMSP and DMS concentrations in pH-stated cultures of the phytoplankton haptophyte Emiliania huxleyi. Four different environmental conditions were tested: ambient, elevated CO2 (+CO2), elevated temperature (+T) and elevated temperature and CO2 (+TCO2). In comparison to the ambient treatment, average DMS production was about 50% lower in the +CO2 treatment. Importantly, temperature had a strong effect on DMS production and the impacts outweighed the effects of a decrease in pH. As a result, the +T and +TCO2 treatments showed significantly higher DMS production of 36.2 ± 2.58 and 31.5 ± 4.66 μmol L−1 cell volume (CV) h−1 in comparison with the +CO2 treatment (14.9 ± 4.20 μmol L−1 CV h−1). As the cultures were aerated with an air/CO2 mixture, DMS was effectively removed from the incubation bottles so that concentration remained relatively low (3.6–6.1 mmol L−1 CV). Intracellular DMSP has been shown to increase in E. huxleyi as a result of elevated temperature and/or elevated CO2 and our results are in agreement with this finding: the ambient and +CO2 treatments showed 125 ± 20.4 and 162 ± 27.7 mmol L−1 CV, whereas +T and +TCO2 showed significantly increased intracellular DMSP concentrations of 195 ± 15.8 and 211 ± 28.2 mmol L−1 CV respectively. Growth was unaffected by the treatments, but cell diameter decreased significantly under elevated temperature. These results indicate that DMS production is sensitive to CO2 and temperature in E. huxleyi. Hence, global environmental change that manifests in ocean acidification and warming may not result in decreased DMS as suggested by earlier studies investigating the effect of elevated CO2 in isolation.