Many lotic fish species use natural patterns of variation in discharge and temperature as spawning cues, and these natural patterns are often altered by river regulation. The effects of spring discharge and water temperature variation on the spawning of shovelnose sturgeon Scaphirhynchus platorynchus have not been well documented. From 2006 through 2009, we had the opportunity to study the effects of experimental discharge levels on shovelnose sturgeon spawning in the lower Marias River, a regulated tributary to the Missouri River in Montana. In 2006, shovelnose sturgeon spawned in the Marias River in conjunction with the ascending, peak (134 m3/s) and descending portions of the spring hydrograph and water temperatures from 16°C to 19°C. In 2008, shovelnose sturgeon spawned in conjunction with the peak (118 m3/s) and descending portions of the spring hydrograph and during a prolonged period of increased discharge (28–39 m3/s), coupled with water temperatures from 11°C to 23°C in the lower Marias River. No evidence of shovelnose sturgeon spawning was documented in the lower Marias River in 2007 or 2009 when discharge remained low (14 and 20 m3/s) despite water temperatures suitable and optimal (12°C-24°C) for shovelnose sturgeon embryo development. A similar relationship between shovelnose sturgeon spawning and discharge was observed in the Teton River. These data suggest that discharge must reach a threshold level (28 m3/s) and should be coupled with water temperatures suitable (12°C-24°C) or optimal (16°C-20°C) for shovelnose sturgeon embryo development to provide a spawning cue for shovelnose sturgeon in the lower Marias River.