The purpose of this study was to examine the relationship between motivation for participation in exercise and the motivational climate in college physical education classes. In Experiment 1, we re-examined two scales in the context of college physical education—the Motivational Climate Questionnaire for physical education classes in Japan (Isogai et al., 2008) and the Motivation Scale for Exercise Adherence, based on self-determination theory (Matsumoto, Takenaka, and Takaie, 2003). By the result of Experiment 1, several items of the motivational climate scale were modified, and the reliability of the two scales were confirmed. In Experiment 2, the relationship between motivation and motivational climate was examined using the two scales. The motivation scale was used before and after physical education classes, whereas the motivational climate scale was used after class. Questionnaire responses showed that intrinsic motivation and identified regulation were maintained at a high level after classes. Additionally, intrinsic motivation and identical regulation were affected before motivation and mastery climate. These results indicate that the relationship between motivation and motivational climate is similar to previous studies. For college students' motivation to increase, the motivational climate in class should be a mastery climate.