A study was conducted under greenhouse conditions on wheat to investigate the utilization of dissolved organic nitrogen (N) in comparison with conventionally applied inorganic N sources (INS). Nitrogen was applied at a rate of 90 kg N ha−1 in an inorganic form, an organic high molecular weight (MW) form (casein, haemoglobin, albumin), and an organic low MW amino acid form (glycine, alanine, valine). Inorganic N sources recorded the maximum response (126% to 150%) in total dry matter (DM) production while dissolved organic nitrogen (DON) sources showed 61% to 116% increase in comparison to the control treatment. Glycine gave the maximum DM production, which was comparable with both INS treatments. In hydroponics, greater utilization occurred and the shoots had a higher N content in comparison to those grown in soil. The concentration of DON and NO3− in soil after wheat harvest was similar in all the treatments.