Spawning site selection by female salmon is based on complex and poorly understood tradeoffs between the homing instinct and the availability of appropriate habitat for successful reproduction. Previous studies have shown that hatchery-origin Chinook salmon (Oncorhynchus tshawytscha) released from different acclimation sites return with varying degrees of fidelity to these areas. To investigate the possibility that homing fidelity is associated with aquatic habitat conditions, we quantified physical habitat throughout 165 km in the upper Yakima River basin (Washington, USA) and mapped redd and carcass locations from 2004 to 2008. Principal components analysis identified differences in substrate, cover, stream width, and gradient among reaches surrounding acclimation sites, and canonical correspondence analysis revealed that these differences in habitat characteristics were associated with spatial patterns of spawning (p < 0.01). These analyses indicated that female salmon may forego spawning near their acclimation area if the surrounding habitat is unsuitable. Evaluating the spatial context of acclimation areas in relation to surrounding habitat may provide essential information for effectively managing supplementation programs and prioritizing restoration actions.