Nitrogen (N) limits plant productivity and its uptake and assimilation may be regulated by N source, N availability, and nitrate reductase activity (NRA). Knowledge of how these factors interact to affect N uptake and assimilation processes in woody angiosperms is limited. We fertilized 1-year-old, half-sib black walnut (Juglans nigra L.) seedlings with ammonium (NH 4 + ) [as (NH 4 ) 2 SO 4 ], nitrate (NO 3 -) (as NaNO 3 ), or a mixed N source (NH 4 NO 3 ) at 0, 800, or 1,600 mg N plant -1 season -1. Two months following final fertilization, growth, in vivo NRA, plant N status, and xylem exudate N composition were assessed. Specific leaf NRA was higher in NO 3 --fed and NH 4 NO 3 -fed plants compared to observed responses in NH 4 + -fed seedlings. Regardless of N source, N addition increased the proportion of amino acids (AA) in xylem exudate, inferring greater NRA in roots, which suggests higher energy cost to plants. Root total NRA was 37% higher in NO 3 --fed than in NH 4 + -fed plants. Exogenous NO 3 -was assimilated in roots or stored, so no difference was observed in NO 3 -levels transported in xylem. Black walnut seedling growth and physiology were generally favored by the mixed N source over NO 3 -or NH 4 + alone, suggesting NH 4 NO 3 is required to maximize productivity in black walnut. Our findings indicate that black walnut seedling responses to N source and level contrast markedly with results noted for woody gymnosperms or herbaceous angiosperms.