Many lichen secondary metabolites have functions related to the environmental conditions of lichen habitats but few studies have compared soil characteristics with lichen species composition or their secondary products. The goal of this study was to investigate the relationship between soil characteristics with lichen species composition and secondary metabolites. Five locations were chosen in Manitoba, each with five sites (transects), and each transect with five quadrats (1 m × 1 m). All species were collected from each of the quadrats, presence of secondary metabolites was determined by thin layer chromatography, and soil characteristics were examined. The results revealed that rounded sand grains were significantly higher in southeastern Manitoba than in other locations, corresponding to a distinct species composition. Angular grains were significantly higher in northern locations, corresponding to a different group of species. Some of the significant relationships between soil characteristics and secondary metabolites include correlations of atranorin with pH, organic matter, and sand content; fumarprotocetraric acid with organic matter and sand content; and usnic acid with pH and organic matter. A better understanding of the role of lichens with respect to soil characteristics will be important for improving soil stabilization in land reclamation. Key words: Cladonia, lichen species, organic matter, sand particles, secondary metabolites.