In the current social, economic, natural and geopolitical context, there is an acute need for research on professional and entrepreneurial skills in line with the challenges and opportunities of the rapidly changing global economy. The aim of this study was to determine the skills required by employees and entrepreneurs in the agri-food sector. This research study consisted of interviews, questionnaires and focus groups on a sample of 111 employers, 288 students and 139 teachers from the North-East Development Region of Romania. The most important skills needed by higher education graduates in the agri-food sector were communication, learning and social skills, with values of 90.5, 74.3 and 70.6 points, respectively. The least appreciated skills were cultural, linguistic and mathematical skills, with values of 17.5, 36.9 and 43.8 points. For developing and running an entrepreneurial activity, the subjects appreciated communication (91.0 points), as in the case of employment-related skills, followed by economic skills (81.4 points) and learning skills (75.4 points). Additional efforts are needed to increase the relevance of practical activities in correlation with the skills required by the market, and it is necessary to strengthen the current partnerships and create new partnerships between universities and the economic environment by employing public and research entities.