This study aimed to examine preservice teachers’ perceptions regarding the use of artificial intelligence (AI) in education. Using convenience sampling, data were collected from 311 undergraduate students at Afyon Kocatepe University, a public university in Türkiye, including 201 students from the Faculty of Sport Sciences and 110 from the Faculty of Education. Participants were enrolled in Primary School Teaching, Physical Education and Sports Teaching, or Coaching Education and Recreation programs with pedagogical formation training. Thus, all participants were preparing for the teaching profession. A quantitative, cross-sectional, comparative survey design was employed. Data were collected using a demographic information form and the 15-item Perception Scale Regarding the Use of Artificial Intelligence in Education, comprising Teaching Perception, Learning Perception, and Ethical Perception subdimensions. The Cronbach’s alpha coefficients were 0.805, 0.805, and 0.609 for these subdimensions, respectively. Skewness values ranged from −0.724 to −0.192 and kurtosis values from −0.579 to 1.552, supporting the use of parametric analyses. Independent-samples t-tests and one-way ANOVA were conducted. Male Faculty of Sport Sciences students scored significantly higher than female students on Teaching Perception. Faculty of Education students who did not use AI scored significantly higher on Ethical Perception than AI users. No other significant gender, age, AI-use, or faculty differences were identified. The findings highlight the importance of promoting pedagogically and ethically appropriate AI integration and strengthening AI literacy in teacher education programs.