This current study was performed to determine the influences of plant spacing, Nitrogen (N) fertilization rate and their effect, on growth traits, yield, and yield components of cotton (Gossypium barbadense L.) cv. Giza 97 during the 2019 and 2020 seasons. A split plot experiment in three replicates was utilized whereas the cotton seeds were planted at 20, 30, and 40 cm, as main plots and nitrogen at 75, 100, and 125%, was in subplots. The results revealed that the planting spacing at 40 cm significantly (p ≤ 0.01) increased plant height, number of fruiting branches per plant, number of bolls per plant, boll weight (BW), lint percentage (L%), seed cotton yield (SCY), lint cotton yield (LCY), seed index and lint index by 165.68 cm, 20.92, 23.93, 3.75 g, 42.01%, 4.24 ton/ha, 5.16 ton/ha, 12.05, 7.86, respectively, as average in both seasons. The application of N fertilizer rate at 125% caused a maximum increase in growth and yield parameters i.e., plant height (169.08 cm), number of vegetative branches (2.67), number of fruiting branches per plant (20.82), number bolls per fruiting branch (1.39), number of bolls per plant (23.73), boll weight (4.1 g), lint percent (41.9%), seed index (11.8 g), and lint index (8.2), while the plants treated with 100% N rates exhibited highest seed cotton yield (4.3 ton/ha) and lint cotton yield (5.6 ton/ha), as average in both seasons. Combining plant spacing at 40 cm between plants with a 100% N fertilizer rate recorded the highest lint cotton yield (5.67 ton/ha), while the highest seed cotton yield (4.43 and 4.50 ton/ha) was obtained from 125% N fertilizer rate under planting spacing 20 and 40 cm, respectively. Conclusively, a wide density (40 cm) with 125% N is a promising option for improved biomass, cotton growth, yield, physiological traits, and fiber quality.