Root plasma membrane H+-ATPase electrochemical equilibrium for optimum coffee plant growth can be modulated by specific ammonium:nitrate (NO3?:NH4+)ratio supply. This study aimed to evaluate the coffee seedlings responses to varying ammonium:nitrate (NO3?:NH4+)ratio and to depict how much NO3? and NH4+ plants can use in terms of growth, nitrogen metabolism, amino acids profile and regulation of root plasma membrane H+-ATPase. Coffee plants were grown in nutrient solution with the following NO3?:NH4+ ratios ( per cent): 100:0; 87.5:12.5; 50:50; 0:100. Plants were grown in nutrient solution for 90 days and evaluated for growth, nitrate reductase activity as well as the modulation of H+-ATPase activity in the plasma membrane of the roots, amino acids profile, chlorophyll a fluorescence parameters and estimated cations and anions taken up by plants. The plants treated with the 87.5:12.5 and 50:50 NO3?:NH4+ ratio showed higher ability to absorb nutrients maintaining balanced uptake and as a consequence, 6 per cent and 29 per cent, the highest dry mass yield as compared to the 0:100 NO3?:NH4+ ratio. In addition, plants supplied with the 87.5:12.5 and 50:50 NO3?:NH4+ ratio had respectively, 58 per cent and 94 per cent, greater photosynthetic capability. Those data suggest that farmers and plant nurseries could implement the 50:50 NO3?:NH4+ ratio of nitrogen sources at coffee plantations and seedlings.