Despite the importance of the theory of evolution (TE) to scientific knowledge, a number of misconceptions continue to be found among biology teachers. In this context, the first objective of this study was to identify the impact of professional development programme (PDP) on teachers’ understanding of nature of science (NOS) and evolution and on the acceptance of this theory. Its second objective was to study the relationship among these variables. Three instruments were used to quantify these variables: the Views of the Nature of Science Version D (VNOS D+), the Assessing Contextual Reasoning about Natural Selection (ACORN), and the Measure of Acceptance of Theory of Evolution (MATE). The results indicate that the PDP had a positive impact on teachers, significantly improving their understanding of the NOS and natural selection, as well as their acceptance of the TE. Furthermore, a positive correlation between the understanding of the NOS obtained by teachers in the first part of the PDP and the understanding and acceptance of evolution that these teachers showed at the end of the programme was determined. However, no relationship between an understanding of the NOS and gains in the understanding and acceptance of evolution was found.