Optimal substrate volumetric water content (θ) and drought tolerance of impatiens, petunia, salvia, and vinca were investigated by growing plants under four constant levels of θ (0.09, 0.15, 0.22, and 0.32 m3·m-3). Gas exchange, quantum efficiency (ΦPSII), electron transport rate (ETR), non-photochemical quenching (NPQ), and leaf water potential (ϒ) were measured for all species, and response of photosynthesis (Pn) to internal CO2 concentration (Ci) was studied in petunia and salvia. Leaf photosynthesis (Pmax) was highest at a θ of 0.22 m3·m-3 for all species and did not differ between a θ of 0.15 and 0.22 m3·m-3 for vinca and petunia. The Pn-Ci response curves for petunia were almost identical at a θ of 0.22 and 0.15 m3·m-3. Regardless of species, ETR and ΦPSII were highest and NPQ was lowest at a θ of 0.22 m3·m-3. Based on these results, a θ of 0.22 m3·m-3 for salvia and impatiens and a slightly lower θ of 0.15 m3·m-3 for vinca and petunia, is optimal. Mean osmotic potential in all treatments was lower in vinca and salvia and resulted in higher turgor potential in these species than other species. Analysis of Pn-Ci response curves indicated that Pn at a θ of 0.09 m3·m-3 was limited by both gas phase (stomatal and boundary layer) and non-gas phase (mesophyll) resistance to CO2 transfer in salvia. At the lowest θ level, Pn in petunia was only limited by gas phase resistance, indicating that absence of mesophyll resistance during drought may play a role in the drought tolerance of petunia.