Rapid and Modular DNA Assembly for Transformation of Marchantia Chloroplasts

The bryophyte Marchantia polymorpha , has attracted significant attention as a powerful experimental system for studying aspects of plant biology including synthetic biology applications. We describe an efficient and simple recursive Type IIS DNA assembly method for the generation of DNA constructs for chloroplast genome manipulation, and an optimized technique for Marchantia chloroplast genome transformation. The utility of the system was demonstrated by the expression of a chloroplast codon-optimized cyan fluorescent protein.