Plant respiratory metabolism is complicated by the fact that the rate of non-photorespiratory mitochondrial CO2 release in the light (Rlight) may be lower than the rate of leaf respiration in the dark (Rdark). A body of work on this topic implies a linkage between light inhibition of respiration and photorespiration, although the direction of effect and underlying mechanisms remain uncertain. In this study we used a variety of short- and long-term environmental manipulations to explicitly manipulate the rate of photorespiration (νo) and quantify the effect on the inhibition of mitochondrial respiration in the light (Rlight:Rdark). We address the following three questions: (i) will the Rlight:Rdark ratio increase or decrease with high CO2 or low O2 and at low temperatures; (ii) does νo correlate with Rlight:Rdark, and if so, in what way; (iii) will suppression of respiration by light (the ‘Kok effect’) be seen to the same extent in Zea mays, a C4 plant, and in Triticum aestivum, a C3 plant? We found that Rlight:Rdark decreased under conditions that suppressed νo in wheat, and this resulted in a positive relationship between Rlight:Rdark and νo. Inhibition of respiration by light in C4 maize did not respond to environmental treatment, and the fixed Rlight:Rdark (0.46–0.72) was consistent with the wheat response, assuming a νo approaching zero. The most likely mechanism to explain this finding is that Rlight increases (or the inhibition of respiration by light decreases) when there is an increase in photorespiration and thus an increase in the demand for TCA cycle substrates associated with the recovery of photorespiratory cycle intermediates in the peroxisome. This work is significant because it combines a comparison of C3 and C4 metabolism with a range of environmental treatments to independently suppress νo.