Little is known about how spatial literacy influences mapping and decision-making related to environmental management and ecosystem services. We assessed the role of spatial landscape literacy in participatory environmental management and ecosystem service assessments in peri-urban Bogota, Colombia. Spatial landscape literacy (SL) was evaluated by testing stakeholder's ability to locate specific landscapes and landmarks. We then assessed if opinions on environmental decisions and ecosystem services were significantly related to SL. We used an online instrument to capture 2,397 respondent's socioeconomic characteristics, SL, ecosystem service perceptions, and opinions concerning relevant environmental issues. We evaluated and measured respondents' self-perceived SL and ability to locate four landscapes in an integrated online map. Positional accuracy was calculated using a Spatial Landscape Literacy Index (SLI). We then tested for effects of socio-demographics on SLI, modeled the relationship between socio-demographics and SL, and tested the relationship between respondents' SL and their opinions on relevant environmental issues and ecosystem services. We found that about 75% of the respondents correctly located 2 of the 4 landscapes. The SLI model was also poorly predicted by socio-demographic variables. However, we found significant relationships between SLI and opinions concerning the environment. No relationship was found between respondents' levels of active participation in local governance and SLI. Overall, SL was little affected by education levels. Participatory processes using maps should ideally measure SL and not assume a priori that participants are spatially literate. Further research is needed to evaluate how spatial technologies and understanding stakeholder's values towards the environment can democratize participatory-based decision-making.