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Plant and Cell Physiology Advance Access originally published online on June 5, 2008
Plant and Cell Physiology 2008 49(7):1084-1091; doi:10.1093/pcp/pcn085
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Agrobacterium-Mediated Transformation of the Haploid Liverwort Marchantia polymorpha L., an Emerging Model for Plant Biology

Kimitsune Ishizaki, Shota Chiyoda, Katsuyuki T. Yamato and Takayuki Kohchi*

Graduate School of Biostudies, Kyoto University, Kyoto, 606-8502 Japan

*Corresponding author: E-mail, tkohchi{at}lif.kyoto-u.ac.jp; Fax, +81-75-753-6127.


  Abstract

Agrobacterium-mediated transformation has not been practical in pteridophytes, bryophytes and algae to date, although it is commonly used in model plants including Arabidopsis and rice. Here we present a rapid Agrobacterium-mediated transformation system for the haploid liverwort Marchantia polymorpha L. using immature thalli developed from spores. Hundreds of hygromycin-resistant plants per sporangium were obtained by co-cultivation of immature thalli with Agrobacterium carrying the binary vector that contains a reporter, the β-glucuronidase (GUS) gene with an intron, and a selection marker, the hygromycin phosphotransferase (hpt) gene. In this system, individual gemmae, which arise asexually from single initial cells, were analyzed as isogenic transformants. GUS activity staining showed that all hygromycin-resistant plants examined expressed the GUS transgene in planta. DNA analyses verified random integration of 1–5 copies of the intact T-DNA between the right and the left borders into the M. polymorpha genome. The efficient and rapid Agrobacterium-mediated transformation of M. polymorpha should provide molecular techniques to facilitate comparative genomics, taking advantage of this unique model plant that retains many features of the common ancestor of land plants.

Keywords: Agrobacterium-mediated transformation - Bryophyte - Hepaticae - Liverwort - Marchantia polymorpha - T-DNA

Abbreviations: GUS, β-glucuronidase; TAIL-PCR, thermal asymmetric interlaced-PCR

(Received March 26, 2008; Accepted May 31, 2008)
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