With about 1000 species in approximately 90 genera, Lejeuneaceae are the largest family of liverworts and make up a large and important part of cryptogamic diversity in the humid tropics. Maximum parsimony, Maximum likelihood and Bayesian analyses of a dataset including four markers (rbcL, psbA, trnL-trnF region of cp DNA, nrITS region) of 134 accessions resulted in similar topologies that support the presence of four main lineages within Lejeuneaceae. Model-based analyses support a division of Lejeuneaceae into two lineages corresponding to the subfamilies Ptychanthoideae and Lejeuneoideae. The latter lineage splits into the tribes Lejeuneeae, Brachiolejeuneeae and the genus Symbiezidium. In contrast, the Maximum parsimony analysis resolves Brachiolejeuneeae and Symbiezidium in serial sister relationships to the remainder of Lejeuneaceae. Sporophyte characters support a split into two subfamilies as seen in the model-based analyses. Some deep nodes remain unresolved, possibly indicating a series of initial diversifications which occurred over a short time period.