The purpose of this research was to examine pre-service science teachers' academic motivation and academic self-efficacy toward biology. The sample consisted of 369 pre-service science teachers who enrolled in the faculty of education of two universities in Turkey. Data were collected through Academic Motivation Scale (AMS) (Glynn & Koballa, 2006) and Academic Self-efficacy Scale (ASES) (Woo, 1999). The findings revealed that pre-service science teachers' academic self-efficacy and academic motivation toward biology are high. Additionally, combination of the independent variables was significantly related to the academic self-efficacy scores, while it was not significantly related to the academic motivation. More specifically, it was found that only university variable made a statistically significant contribution to the prediction of pre-service science teachers' academic motivation and academic self-efficacy. There were strong, positive correlations between pre-service science teachers' academic motivation and academic self-efficacy and subdimensions of AMS and ASES except "test anxiety". The relationship of test anxiety with all of the subdimensions was small.