Agrobacterium mediated transformation of Marchantia spores v4

This method produces random integration of DNA into the genome of Marchantia. The technique relies on the capacity of Agrobacterium tumefasciens to transfer DNA to plants. This DNA can be specified as it is flanked by sequences of DNA called the Right and Left Border (RB & LB), named as a whole as the T-DNA. The original tumour-inducing (Ti) plasmid from Agrobacterium was adapted to a binary plasmid system allowing easy cloning of the T-DNA part of the system in E. coli before being electroplated into Agrobacterium and then transfection into plants by infection. This protocol is based onIshizaki et al., 2008, optimised in Jim Haseloff lab,Plant Sciences, University of Cambridge, by Nuri Purswani and Bernardo Pollak. At the moment Linda Silvestri and Susana Sauret-Gueto are seeing on further optimisation, including feedback from other researchers working in Marchantia, like Bo Xu.