<p>Background: In 2015, the Ministry of Education, Culture, Sports, Science, and Technology （MEXT） published a report on sexual minority students, including those with gender identity disorder. This promoted discussions on sexual minorities in schools. Reports showed that teachers did not respond to such students adequately, and teachers may hurt the feelings of sexual minority students, suggesting that school is not a safe space for them. In particular, support for sexual minority students in physical education classes is important as shown in the MEXT report.</p><p>Objective: This study aimed to investigate and identify the main characteristics of the knowledge and attitudes of school teacher trainees who majored in health and physical education （HPE） toward sexual minorities.</p><p>Methods: A questionnaire survey was administered to 393 students belonging to "A" university for training students to be teachers. There were 377 questionnaires recovered and 351 valid respondents （response rate ＝89.3%）, of which 127 students majored in HPE, and 224 students majored in other subjects. The survey included basic information regarding respondents, learning experiences and knowledge about sexual minorities, attitudes toward sexual minorities, including homophobia （irrational fear and hatred of homosexuals）, transphobia （irrational fear and hatred of transgender individuals）, seeing and hearing aggression toward sexual minorities, experiences involving sexual minorities, and perspectives on education.</p><p>Results: In the group of students who majored in HPE, "normative-men" （cis-gendered, heterosexual） tended to make sexual minorities invisible and reinforce homophobia and transphobia more than "normativewomen" （cis-gendered, heterosexual）. In the group of "normative-men, "students who majored in HPE showed this tendency more than students who majored in other subjects. Students who were not close to sexual minorities tended to make sexual minorities invisible and reinforce homophobia and transphobia more than students who were close to them.</p><p>Conclusion: This study showed that the students who majored in HPE demonstrated strong homophobia and transphobia more than the students majoring in other subjects.</p><p> However, many of students who recognized the existence of sexual minorities had known others who had come out or disclosed their sexual identity; the homophobia and transphobia of such students subsequently weakened. Thus, the act of coming out is a key factor in encouraging awareness of the existence of sexual minorities.</p>