Agroecology represents a pertinent option to improve the sustainability of agriculture. To promote its application, agroecological concepts should be taught to students and professionals in the agricultural sector. However, most agricultural courses are not adapted to teach these concepts due to little interactivity or interdisciplinarity, and a lack of a systems approach to farm management. Serious games help to fill these gaps by simulating complex models in which players can learn by doing. We thus developed a serious computer game, called SEGAE (SErious Game for AgroEcology learning), which represents a mixed crop-livestock farm and assesses impacts of farming practices on indicators related to environmental, economic, and social sustainability. Its pedagogical interest was evaluated through two types of surveys given to university students who played the game during a one-week workshop: A knowledge survey on agroecology, and a feedback survey based on flow theory. Results showed that students increased their knowledge of agroecology significantly, particularly those who had had little knowledge of crop production. More than 86% of the students enjoyed the game, appreciating its interaction and feedback. Thus, SEGAE is an interesting tool to help students acquire knowledge of agroecology in a fun way by facilitating interdisciplinary and collaborative learning.