Development of molecular genetic resources for the liverwort, Marchantia polymorpha II <Simple and efficient Agrobacterium-mediated transformation>

Abstract

Here we report a simple and efficient transformation of the liverwort Marchantia polymorpha using Agrobacterium tumefaciens. Hundreds of transformants per single sporangium were obtained by co-cultivation of young gametophytes with A. tumefaciens strain GV2260 carrying the binary vector pIG121-Hm that contains the β-glucronidase (GUS) gene with an intron, and a selective marker, the hygromycin phosphotransferase (hpt) gene. GUS activity staining showed that all the transformants examined expressed the foreign gene. Southern blot analyses confirmed the integration of 1~5 copies of the hpt gene randomly into the genome. It took only 3 to 4 weeks to establish individual transgenic plants after starting suspension culture of gametophytes. The efficient and rapid Agrobacterium-mediated transformation protocol can be applied not only to introduction of constructs of interest, but also to T-DNA insertional mutagenesis in M. polymorpha.