Nocturnal root and meristem temperature (RMT) can have a strong effect on rice growth and yield. However, underlying mechanisms are not well understood. To investigate the effects of night-time RMT on photosynthesis biomass allocation and activities of antioxidant enzymes, we designed a hydroponic system that maintained the following daily patterns of day/night temperature: 18/28 °C (HNT) or 28/18 °C (LNT). Rice plants cv. IR64 were grown in the greenhouse and subjected to either HNT or LNT. HNT stimulated growth and tillering but did not affect biomass allocation. HNT plants increased total biomass by 16 and 35%, depending on time of exposure. HNT increased rates of photosynthesis (Pn) compared to LNT plants in leaves of different ages. Overnight carbohydrate remobilisation was larger in HNT than in LNT plants, particularly at 16 days after treatment (dat), when Pn and relative growth rates were highest. Leaf soluble protein concentrations and specific leaf area were not affected by RMT, indicating higher photosynthetic nitrogen use efficiency in HNT plants. Super Oxide Dismutase, Ascorbate Peroxidase, and Glutathione Reductase activities did not respond to RMT, indicating no change in the production of reactive oxygen species in LNT plants despite lower photosynthesis rates. HNT increased sink demand by stimulating tillering, the increased sink demand upregulated the source activity through a larger leaf area per plant and a higher Pn throughout the canopy. The hydroponic system described here was able to control the temperature of the nutrient solution effectively, the installation of a second pump directly circulating the nutrient solution from and back to the reservoir through the cooling system allowed reaching the target temperature within 1 h. This system opens new opportunities to characterise plant responses to RMT alone or in combination with other environmental drivers.