The understanding of real-world application of scientific knowledge such as the evolutionary theory has been emphasized in science curriculum. Despite this, many students and biology educators misunderstand the use and practical application of the evolution theory. The purpose of this study is to explore variables related to the understanding of practical application of evolution. Methodologically, we employed large-scale survey that examined three independent variables (major, academic year, and religion) and five dependent variables (understanding of practical application of evolution, knowledge of, interest in, and acceptance of evolution, and religiosity). Two hundred and thirty three biology pre-service teachers and 405 non-biology pre-service teachers participated in the survey. The results demonstrate that the biology teacher education program does not help in increasing the understanding of the practical application of evolution. Rather, the understanding of practical application of evolution was associated with their religion. In addition, the understanding of the practical application of evolution was associated with the acceptance of and interest in evolution. The results of this study inform the development of new biology teacher education programs to improve their understanding of practical application of the evolution theory. We also suggest further areas of research for other scientific concepts that could be used to gauge teachers` understanding of the practical application of scientific knowledge.