Background: This study investigated the effect of an adapted physical activity (APA) course on knowledge and perceptions of preservice trainees regarding physical activity (PA) and autism spectrum disorder in 3 areas: knowledge/perspectives, importance and ease of improving developmental domains, and importance and ease of improving motor skills. Methods: Four hundred upper-level undergraduate students were recruited to participate in this survey-based study (251 APA students and 149 non-APA students participated). Survey data were analyzed using multivariate analyses of variance. Results: Participants estimated that the moderate to vigorous PA recommendations are 39.34 minutes per day, that 46.65% of moderate to vigorous PA occurs during school, and that 61.03% of children have motor difficulties. Participants perceived activities of daily living, sleep habits, and heart health as the easiest domains to improve, and problem behaviors, social skills, and self-esteem as the most difficult domains to improve. Knowledge/perspectives regarding autism spectrum disorder and PA were different by APA exposure (F12,324 = 3.11, P < .001). Differences included self-efficacy in providing PA advice, knowledge of PA guidelines, and willingness to provide motor assessment referrals. Students differed by APA exposure in the importance of developmental domains (F8,381 = 4.37, P < .001) but not ease of improving those domains. Conclusion: Results suggest that APA education and contact with children with disabilities improves self-efficacy, perspectives, and knowledge of PA and motor concerns in children with autism spectrum disorder.