This study was conducted during the 2020 and 2021 growing seasons to evaluate the effects of spraying various growth regulators (Volex 250 ppm, Cytoking 250 ppm, Cytokinin 250 ppm, and Gibberellin 250 ppm) on the morphological, physiological, and yield characteristics, as well as the fruit quality of Grande Naine banana plants (Musa cavendishii) grown in Sohag, Egypt. The results showed significant improvements in pseudostem height, circumference, and the number of leaves per plant in treated plants compared to the control. The most notable increases in yield components, including the number of fingers and hands per bunch, hand weight, and bunch weight, were recorded in plants treated with Cytokinin 250 ppm, which showed statistically significant improvements over the control (p < 0.05). In terms of fruit quality, treatments with Volex and Cytoking at 250 ppm resulted in higher total soluble solids (TSS) and TSS/acid ratio, while lowering fruit acidity compared to untreated plants. Based on these findings, we recommend the use of Cytokinin 250 ppm to enhance banana yield and fruit quality under the environmental conditions of Upper Egypt.