The aim of this study is to examine the relationship between the psychological needs of athletes who attend gyms, the spirituality index of well-being, and motivation in sports, and to determine the mediating role of the spirituality index of well-being between psychological needs in sports and motivation in sports. Although sports spirituality is rapidly developing in the international literature, sports and spirituality have not been studied together in the national literature before. Therefore, this study holds the distinction of being one of the pioneering research efforts on the subject of spirituality in athletes in Türkiye. In this context, the data of 422 athletes, 176 women and 246 men, who have been attending gym facilities in Antalya for at least one year, were included in the research. Participants were administered a personal information form, Psychological Need States. It was found that the spirituality index of well-being is positively related to all satisfaction subscales and negatively related to all frustration subscales of psychological needs and that the life scheme is related to many subscales of sport motivation. Furthermore, it was also found that the spirituality index of well-being is a mediating variable between psychological needs in sports and motivation in sports. The data draw attention to the importance of sports spirituality. It is recommended to integrate spirituality into healthcare for athletes.