The aim of this paper is to investigate if stakeholder participation increases the legitimacy of nature reserves in the surrounding community. Most previous studies of the effects of stakeholder participation in natural resource management have relied on case studies, but in this paper we use a combination of panel data from a two-wave survey (2008 and 2013) of 92 Biosphere Reserves (BRs) in 36 countries and semi-structured interview data from 65 stakeholder respondents in a sub-sample of 10 BRs to systematically investigate the effects of stakeholder participation on the legitimacy of the natural reserve in the local community. The data cover four levels of stakeholder participation: (1) Information, (2) Implementation, (3) Involvement and (4) Representation. These levels roughly correspond to rungs on Arnstein’s ladder of participation, and the expected outcome is that the legitimacy of the nature reserve will increase in the surrounding local community as the degree of participation increases. However, findings suggest that there is no linear relationship between participation and legitimacy: climbing upwards on Arnstein’s ladder of participation does not uniformly enhance the level of legitimacy of the nature reserve in the local community. Instead, a practice-based form of participation is what seems to increase legitimacy.