Agartrap protocols on your benchtop: Simple methods for agrobacterium-mediated genetic transformation of the liverwort marchantia polymorpha

Abstract

Agrobacterium-mediated genetic transformation is a powerful technique in plant biology. We recently developed a simplified Agrobacterium-mediated genetic transformation method for the liverwort Marchantia polymorpha, named AgarTrap (agar-utilized transformation with pouring solutions). AgarTrap is easy to perform; all procedures can be completed within a week using a single plate of solid medium, and basic operations involve simply pouring the appropriate solutions onto the solid medium. Thus far, we have developed three types of AgarTrap methods (S-AgarTrap, G-AgarTrap, and T-AgarTrap) using three different M. polymorpha tissues: sporelings, intact gemmalings, and mature thallus pieces, respectively. Each AgarTrap method can be used to transform tissues at high efficiency, thereby producing sufficient numbers of transformants for study. The ease and efficiency of these AgarTrap methods will likely prompt widespread molecular biological analyses of M. polymorpha. In this review, we describe the basic characteristics of the three AgarTrap methods and present the detailed protocols used in our laboratory.