Variations to the global wetland CH4 source strength in response to changes in orbital insolation patterns and atmospheric CO2 concentration ([CO2]a) are hypothesized to play an important role in determining glacial-interglacial variations in atmospheric CH4 concentration ([CH4]a). Here the interactive effects of temperature, a major controlling variable determining wetland CH4 flux, and the low [CO2]a of glacial intervals are investigated for the first time. We measured the temperature dependence of CH4 emissions from replicated mesocosms (n = 8 per CO2 treatment) collected from a minerotrophic fen and an ombrotrophic bog incubated in either ambient (c. 400 ppm) or glacial (c. 200 ppm) [CO2]a located in the United Kingdom. CH4 fluxes were measured at 5°C, 10°C, 15°C, 20°C, and 25°C and then in reverse order over a 20 day period under each [CO2]a treatment. Results showed that the Q10 temperature response of CH4 emissions from the Carex/Juncus-dominated fen declined significantly by approximately 39% under glacial [CO2]a (ambient [CO2]a = 2.60, glacial [CO2]a = 1.60; P < 0.01). By contrast, the response of CH4 emissions from the Sphagnum-dominated bog remained unaltered (ambient [CO2]a = 3.67, glacial [CO2]a = 3.67; P > 0.05). This contrasting response may be linked to differences in plant species assemblage and the varying impact of CO2 starvation on plant productivity and carbon availability in the rhizosphere. Furthermore, our results provide empirical evidence to support recent model-based indications that glacial-interglacial variations in [CH4]a may be explained by changes in wetland CH4 source strength in response to orbitally forced changes in climate and [CO2]a.