Egg sampling confirmed that Suwannee River Gulf sturgeonAcipenser oxyrinchus desotoi, a subspecies of Atlantic sturgeonA. o. oxyrinchususe the same spawning site at river kilometer (rkm) 215 from the mouth of the river each year. Forty-nine eggs were recorded in 1995, and 368 were recorded in 1996. Spawning began 4–7 d after the March new moon in both years and lasted 10–11 d; in 1996, a second 10-d spawning round began on the April new moon. Developmental synchrony among eggs recovered suggested several discrete spawning events in both years. Total eggs deposited for three 1996 sampling days was estimated as 405,600–711,000/d, approximating the fecundity range of a large female Gulf sturgeon. Eggs were found only in the southern half of the river, an area with surface currents of 0.5–1.5 m/s and numerous eddies producing reverse bottom currents of 0.1–0.5 m/s. Egg substrate consisted of bedrock limestone thinly overlain by fine sand and densely distributed elliptical gravel 2–10 cm in diameter. Eggs were found predominantly in depths of 2–4 m at water temperatures of 17–21°C, conductivities of 50–100 μS, and dissolved oxygen levels exceeding 5.0 mg/L. The Cody Scarp, 15 rkm above the spawning ground, may mark the upstream limit of spawning areas in the river. Three 2–4-month-old riverine juveniles (82–115 mm total length, TL) collected are the smallest yet captured from any river. Data for 18 riverine age-0 juveniles (to 350 mm TL) suggest that this stage lasts 6–10 months, terminating with migration of fish to the river mouth in January–February. Less than 2% of 461 juveniles captured at the estuarine river mouth (1990–1993) were under 350 mm TL. Riverine age-0 fish were collected over long shallow stretches (typically <4 m deep) of relatively barren sand (rkm 12–238).