The dioecious liverwort, Marchantia polymorpha L., is an emerging model plant. Various molecular biological techniques have been optimized for M. polymorpha for the past several years, and recently we reported a simplified Agrobacterium-mediated transformation method using sporelings (immature thalli from spores) of M. polymorpha. This method, termed AgarTrap (Agar-utilized Transformation with Pouring Solutions), completed by exchanging appropriate solutions on a single Petri dish to produce a sufficient number of independent transgenic sporelings. However, because spores are produced by crosses between males and females, the genetic backgrounds of resulting transgenic sporelings are not uniform. To easily produce transgenic liverworts with a uniform genetic background using AgarTrap, we developed an AgarTrap-mediated transformation method using intact gemmae/gemmalings produced by asexual reproduction. Using AgarTrap with male and female gemmae/gemmalings produced a sufficient number of independent transgenic gemmalings with uniform genetic backgrounds. The optimized transformation efficiencies were approximately 30 and 50 % in males and females, respectively. As with AgarTrap using sporelings, AgarTrap using intact gemmae/gemmalings will be useful in promoting studies of the molecular biology of M. polymorpha.
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The authors thank Dr. Christopher Suarez (University of Notre Dame) for critical reading of the manuscript. The authors also thank Dr. Takayuki Kohchi (Kyoto University) for providing Tak-1 and BC3-38 lines. This work was supported by the Plant Transgenic Design Initiative of University of Tsukuba (Y. K.), the Japan Society for the Promotion of Science Research (JSPS) KAKENHI (No. 26840088) (Y. K.) and research Projects [CORE Adopted, CDI-F, UU-COE and UU-COE-next] of Utsunomiya University (Y. K).
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