Homophobia and transphobia are feelings of rejection and hostility towards homosexual and transgender people, which arise from prejudices and fears towards sexual diversity. The objective of the present research was to compare the homophobia and transphobia levels of physical education students and other pedagogies in Chile. To this end, the homophobia scale EHF-10 and the negative attitudes to trans people scale were applied to 384 students of 15 pedagogies from a university in Chile. The results show that homophobia and transphobia scores tend to be low in students of all careers. However, physical education students have the highest scores among all/future teachers, while biology, Spanish, English, history and geography students/students, Basic education and differential education have the lowest scores on one or both scales. It is concluded that there are differences in the levels of homophobia and transphobia according to the course, with the highest rates in physical education, although the scores of both scales are low.