The values (i.e., importance) that people place on ecosystems have been identified as a crucial dimension of sustainable management of social-ecological systems. Recently, the call for integrating plural values of ecosystems beyond intrinsic and instrumental values has prompted the notion of “relational values.” With the aim of contributing to environmental management, we assess the environmental motivations (i.e., egoistic, biospheric, altruistic) and values that people attribute to the ecosystems of the mid-upper stream of the Otún River watershed, central Andes, Colombia. We analyzed 589 questionnaires that were collected in urban and rural areas of the Otún River watershed using the nonparametric Mann-Whitney U test and logistic regressions. We found salient biospheric motivations and the attribution of plural values (i.e., intrinsic, relational, and instrumental) to the ecosystems of the mid-upper stream of the Otún River watershed. Particularly, relational values were the most frequently mentioned value domain. Further, our results showed that environmental motivations and socioeconomic factors are associated with the expression of different value domains. We found negative associations between egoistic motivations and intrinsic values and between rural respondents and instrumental values. We found positive associations between altruistic motivations and relational values and between rural respondents and both intrinsic and relational values. In light of our results, we argue that intrinsic, instrumental, and relational values coexist in people’s narratives about the importance of ecosystems. Plural valuation approaches could be enhanced by differentiating relational from instrumental values and by expressing them in nonmonetary terms. We argue that multiple values of ecosystems expressed by rural and urban societies should be included in environmental management to tackle social conflicts and consider the diverse needs and interests of different social actors.