This comprehensive review focuses on the role of bio-regulators in the growth and development of fruit and vegetable crops. Bio-regulators, also known as plant growth regulators or plant hormones, play a crucial role in regulating various physiological processes in plants, including cell division, elongation, differentiation, and fruit ripening. The review explores the major types of bio-regulators commonly used in fruit and vegetable cultivation, including auxins, gibberellins, cytokinins, abscisic acid, and ethylene. It examines their functions and effects on plant growth, flowering, fruit set, fruit development, and post-harvest characteristics. The review also discusses the widespread application of bio-regulators in horticultural practices, including seed treatment, foliar application, and post-harvest treatments. It highlights the potential benefits of using bio-regulators, such as improving crop yield, enhancing fruit quality traits, delaying senescence, and reducing post-harvest losses. Moreover, the review delves into the mechanisms of action of bio-regulators at the molecular, physiological, and biochemical levels. It investigates their interactions with various plant growth processes, signaling pathways, and gene expression patterns. Furthermore, the review addresses the challenges and limitations associated with the use of bio-regulators in fruit and vegetable production, such as dosage optimization, application timing, and potential negative impacts on the environment.