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Liverworts are key species for studies of plant evolution, occupying a basal position among the land plants.  Marchantia polymorpha  has emerged as a highly studied model liverwort, and many relevant techniques, including genetic transformation, have been established for this species.  Agrobacterium -mediated transformation is widely used in many plant species because of its low cost. Recently, we developed a simplified  Agrobacterium -mediated method for transforming  M. polymorpha , known as AgarTrap ( agar -utilized  tra nsformation with pouring solutions). The AgarTrap procedure, which involves culturing the liverwort tissue in various solutions on a single solid medium, yields up to a hundred independent transformants. AgarTrap is a simple procedure, requiring minimal expertise, cost, and time. Here, we investigated four factors that influence AgarTrap transformation efficiency: (1) humidity, (2) surfactant in the transformation buffer, (3)  Agrobacterium  strain, and (4) light/dark condition. We adapted the AgarTrap protocol for transforming intact gemmalings, achieving an exceptionally high transformation efficiency of 97%. The improved AgarTrap method will enhance the molecular biological study of  M. polymorpha . Furthermore, this method provides new possibilities for improving transformation techniques for a variety of plant species.

Improved G-AgarTrap: A highly efficient transformation method for intact gemmalings of the liverwort Marchantia polymorpha