Distinct distribution patterns were observed for several species of charophytes during a survey in Chile. We attempted to link these patterns to climatic conditions, but no direct causality between specific climatic factors and charophyte growth conditions could be identified. In this paper, correlations between water chemistry and occurrence of charophytes were tested as an alternative approach to explain the distribution pattern. Datasets with information about conductivity, alkalinity, pH and concentrations of calcium, magnesium, sodium, potassium, sulphate and chloride from sites inhabited by at least one charophyte species were analysed by principal component analysis. The results show that the distribution patterns of at least some species can be explained by niche differences with respect to water chemistry. The genera Chara and Nitella are in general well separated by their preferences for a given pH, alkalinity and conductivity range. Within the genus Chara, Chara squamosa, Chara braunii, Chara fulgens, and a yet unidentified taxon Chara sp., differ from this genus-specific pattern with respect to at least one parameter, whereas Chara vulgaris occupies a broad niche, covering almost the full range of all parameters except conductivity. Within the genus Nitella, niche separation by water chemistry was only possible between Nitella tenuissima and Nitella asagrayana. The chemical niche of Chara braunii was more closely related to the niche of the genus Nitella than the niche of other Chara species.