Legume-based cropping systems have the potential to internally regulate N cycling due to the suppressive effect of soil N availability on biological nitrogen fixation. We used a gradient of endogenous soil N levels resulting from different management legacies and soil textures to investigate the effects of soil organic matter dynamics and N availability on soybean (Glycine max) N 2 fixation. Soybean N 2 fixation was estimated on 13 grain farm fields in central New York State by the 15 N natural abundance method using a non-nodulating soybean reference. A range of soil N fractions were measured to span the continuum from labile to more recalcitrant N pools. Soybean reliance on N 2 fixation ranged from 36% to 82% and total N 2 fixed in aboveground biomass ranged from 40 to 224 kg N ha −1 . Soil N pools were consistently inversely correlated with % N from fixation and the correlation was statistically significant for inorganic N and occluded particulate organic matter N. However, we also found that soil N uptake by N 2 -fixing soybeans relative to the non-nodulating isoline increased as soil N decreased, suggesting that N 2 fixation increased soil N scavenging in low fertility fields. We found weak evidence for internal regulation of N 2 fixation, because the inhibitory effects of soil N availability were secondary to the environmental and site characteristics, such as soil texture and corresponding soil characteristics that vary with texture, which affected soybean biomass, total N 2 fixation, and net N balance.