Complete nuclear-encoded (18S) small subunit rRNA gene sequences were determined for four charophycean green algae, Chlorokybus atmophyticus, Coleochaete orbicularis, Klebsormidium flaccidum, and Nitella sp. Chlorokybus atmophyticus and Coleochaete orbicularis have been previously suggested to represent the most basal and most derived taxa within the charophytes, respectively. However, parsimony analysis of our 18S rDNA sequences along with a selection of other complete green algal and land plant 18S rDNA sequences yields a gene tree topology in which Chlorokybus is the most basal taxon, followed by the branching of Coleochaete and Klebsormidium. Two ''sister'' clades then diverge, one including Nitella and the land plants, and the second. members of the Chlorophyceae and Pleurastrophyceae. Despite producing slightly different gene tree topologies than those inferred from parsimony, distance analyses of the 18S rDNA sequences also do not indicate a strong affinity between the land plants and Cleochaete. Rather, Klebsormidium and Coleochaete are virtually equidistant from the land plant taxa. Other data are needed in order to assess the unexpected findings reported here, particularly the position of Coleochaete.