Nodulated plants of six varieties of Stylosanthes were grown at a constant temperature of 30°C for 21 days in a controlled environment and then for a further 21 days at a range of day/night root temperatures. Dry matter and nitrogen content of S. hamata cv. Verano and CPI40264A were measured after growth at day/night root temperatures of 30/30, 40/30, 50/30, 60/30 and 60/35°C. Similarly, growth and nitrogen content were measured for Verano, CPI40264A, S. gvianensis cv. Oxley and cv. Cook, S. humilis cv. Lawson and S. scabra cv. Seca after growth at root temperatures of 25/20, 25/25, 30/25, 30/30, 35/30 and 40/30°C. There were strong interactions of root temperature with variety and harvest time (31 and 42 days) but all varieties grew well even at the highest temperatures (50–60°C) provided they were supplied with nitrogen. Symbiotically dependent plants of S. hamata cv. Verano were more tolerant of high root temperatures and maintained better growth and nitrogen fixation at 40/30°C than those of CPI40264A, especially after the 31 day harvest. All six varieties responded similarly, but to differing extents, to the lower range of root temperatures. Nitrogen fixation was more severely affected than dry weight by high root temperature.