In modern oenology, supplementation of nitrogen sources is an important strategy to prevent sluggish or stuck fermentation. The present study thoroughly determined the effect of nitrogen addition timing and nitrogen source type on fermentation kinetics and aroma production, carried out by yeast strains with low and high nitrogen requirements. The results revealed that yeast strains with different nitrogen requirements have divergent reactions to nitrogen addition. Nitrogen addition clearly shortened the fermentation duration, especially for the high-nitrogen-demanding yeast strain. Nitrogen addition at 1/3 fermentation was the most effective in terms of fermentation activity, nitrogen assimilation, and production of acetate esters. Interestingly enough, yeast cells preferentially took up amino acids related to fermentative aroma synthesis with the addition at 2/3 fermentation. The addition of nitrogen sources also largely affected the production of important metabolites. Generally speaking, acetic acid, glycerol, and succinic acid reduced with the supplementation of nitrogen sources. The results revealed significant application importance for the winemaking industry.