The current turbulence in the economic environment has led to the need for new managerial tools to be used such as stakeholder analysis. Since there are a large number of stakeholder groups in small and medium-sized enterprises, the identification and prioritisation of stakeholders (as a key part of stakeholder analysis) is considered an important element in their management. The purpose of this article is to identify, prioritize and map key stakeholder groups in small and medium-sized enterprises in the Czech Republic, with a view to being able to generate stakeholder profiles of the most important stakeholder groups. The identification of the key stakeholder groups was based on research conducted by the author, whilst the prioritisation of the stakeholder groups on the basis of the professional judgement of the respondents. A power-interest matrix was subsequently compiled to map the stakeholder groups. The results of the research identified customers, employees, competitors, suppliers and media as being the five key stakeholder groups in small and medium-sized enterprises.