A study was conducted to define culture conditions for in vitro growth arbuscular mycorrhizal fungi (AMF) with liverworts as hosts. Lunularia cruciata (L.) Dumortier ex. Lindberg developed in vitro monoxenic mycothalli with both Glomus proliferum Dalpé & Declerck (MUCL 41827) and Glomus intraradices Schenck & Smith (MUCL 43204). AMF inoculated plants were co-cultured in plastic Petri dishes with semi-solidified medium supplemented with sucrose and grown under filtered light. Mycothalli of L. cruciata produced external hyphae and spores in quantities equivalent to those obtained with Ri T-DNA transformed root systems.