This protocol describes how to transform moss (Physcomitrella patens) protoplasts using polyethylene glycol (PEG)-mediated DNA uptake. The transformation rates for direct uptake by protoplasts of DNA with and without genomic sequence (a targeting construct) are typically 10 and 10, respectively. (These are the frequencies of stable transformants among regenerants surviving the transformation pr...

The potential role of soil fungi in the invasion of the Brazilian pepper tree (Schinus terebinthifolius-BP) in Florida is not known; although the low biotic resistance of Florida soils is often invoked to explain the prevalence of many invasive species. To gain an initial insight into BP's mycorrhizal associations, this study examined the rhizobiome of BP and two native plants (Hamelia patens a...

This article includes a series of methods for culturing the moss Physcomitrella patens at all stages of its life cycle. Gametophytes are axenically cultured on solid agar-based media and in shaken liquid cultures. For long-term storage of gametophytes, cultures are maintained on solid medium at 10°C in a very short day. Cryopreservation may also be used. Finally, sporophytes are generated by se...

To regenerate auxotrophic mutants of Physcomitrella patens, two media of increasing complexity were developed. The survival rate of protoplasts was around 30% higher on full medium when compared to standard minimal medium. Protoplast survival was higher in a medium containing 2.5 mmol/L ammonium tartrate compared to a medium with 5 mmol/L of this compound. Solid medium had a positive effect on ...