Prickly pear (Opuntia ficus-indica (L.) Mill. 1768) is cultivated in several dry and semi-dry areas of the world to produce fresh fruit, bioenergy, cosmetics, medicine, and forage. One of the main production constraints is the presence of many seeds within the fruit, which can negatively influence both the fresh-fruit market price and industrial transformation processes. In this study, different gibberellic acid (GA₃) concentrations were tested for their ability to produce well-formed and seedless fruits. Different application methods (injection and spraying) and concentrations of GA₃ (0, 100, 200, 250, and 500 ppm) combined with floral-bud emasculation were applied to a commercial plantation in southern Italy to evaluate their effects on the weight, length, and diameter of the fruits, total seed number, hard-coated viable seed number, and seed weight per fruit. The results indicated that the application of 500 ppm GA₃ sprayed on emasculated floral buds was the most effective method for reducing seed numbers of prickly pear fruits (−46.0%). The injection method resulted in a very low number of seeds (−50.7%) but produced unmarketable fruit. Observed trends suggest the need to investigate the impact of higher GA₃ concentrations and the applicability of a maximum threshold. Further studies are needed to increase our understanding of the physiological effects of the gibberellic acid pathway through productive tissue in terms of organoleptic and fruit quality.