We describe physical and environmental characteristics associated with Micropterus notius (Suwannee Bass) spawning in the Ichetucknee River, FL. We located 24 Suwannee Bass nests from March to June (75% found May and June). Nests were found in relatively shallow waters (mean = 95 cm) with substrates dominated by organic material (63%). Suwannee Bass nests were located near cover, in low-flow areas (mean velocity = 0.01 m/s), and at a mean temperature of 21.7 °C. We did not detect any relationships from regressions between the frequency of nesting and temperature, photoperiod, river stage, discharge or lunar cycle. Ninety-two percent of nests contained submerged aquatic vegetation (SAV), which are likely important to nesting areas as stream-velocity refuges that reduce sediment wash-in and egg disturbance during spawning. Descriptions of spawning habitat of this range-restricted species have implications for conservation and management decisions.