The response of potato (Solanum tuberosum L. 'Superior’) to glyphosate [N-(phosphonomethyl)glycine] at rates between 0.28 and 2.24 kg ae/ha was determined in four environments: 24/13 or 13/4 C temperature regimes with light levels of 650 ± 53 or 320 ± 45 μE · m−2 · S−1. Light level did not affect phytotoxicity ratings or shoot fresh weight following glyphosate application. Greater phytotoxicity and reduced fresh weight accumulation were observed from glyphosate at rates above 0.56 kg/ha in a high-temperature regime than for comparable plants grown at low temperature. Studies of 14C-glyphosate uptake and translocation indicated that the differences in phytotoxicity were due to less glyphosate absorption at low temperature and not a shift in distribution patterns within the plant.