This research presents a case study on the knowledge of pre-service teachers of the school of lower secondary education on biodiversity conservation in Rwanda. It critically examines the implication of the level of knowledge on attitudes and behaviors towards biodiversity conservation and the potential implications of a lack of the courses focusing on biodiversity conservation in the school of lower secondary education, and presents empirical data from a survey and group discussions. Results showed that generally the level of knowledge of pre-service teachers on biodiversity conservation is high, and there is positive correlation between knowledge, awareness and attitude, which in turn may contribute to sustainable biodiversity conservation in Rwanda. Research concluded that there should be another research on the side of primary and secondary students in order to verify if the skills of teachers in biodiversity conservation are fully taught to the students.