Bryophytes, Physcomitrella patens, Marchantia polymorpha, in vitro cultivation
Standardized in vitro cultivation of plant model organisms is a powerful tool for modern plant science, ranging from evo-devo or physiological studies to functional genomics and biotechnology. Bryophytes (comprising liverworts, mosses, and hornworts) are representatives of early diverging land plants and share a unique life strategy, with the haploid gametophyte being the dominant generation. Next to their interesting phylogenetic position and physiology (e.g., desiccation tolerance) they possess large varieties of metabolites and other biologically active compounds, which provide a high potential for applied as well as basic research. In vitro cultivation of bryophytes serves as basis for investigations on these plants under highly standardized conditions, and potentially facilitates an ex situ conservation of endangered species. Here, we describe protocols for axenic in vitro cultivation of mosses and liverworts, starting from non-axenic material collected in the field. The protocols include spore and thallus sterilization procedures, media preparation, and cultivation under standardized conditions. These methods can be used to establish further bryophyte model organisms for basic and applied research.