Abstract
We describe a simple and efficient plastid transformation method for the liverwort, Marchantia polymorpha L. Use of rapidly proliferating cells such as sporelings, which are immature thalli developing from spores, as targets made plastid transformation by particle bombardment efficient. Selection on a sucrose-free medium and linearization of the transformation vector significantly improved the recovery rate of plastid transformants. With the method described here, a few plastid transformants are obtained from a single bombardment of sporelings. Homoplasmic transformants of thalli are obtained immediately after primary selection.
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References
Shimamura M (2016) Marchantia polymorpha: Taxonomy, phylogeny and morphology of a model system. Plant Cell Physiol 57:230–256
Bowman JL, Kohchi T, Yamato KT, Jenkins J, Shu S, Ishizaki K, Yamaoka S, Nishihama R, Nakamura Y, Berger F, Adam C, Aki SS, Althoff F, Araki T, Arteaga-Vazquez MA, Balasubrmanian S, Barry K, Bauer D, Boehm CR, Briginshaw L, Caballero-Perez J, Catarino B, Chen F, Chiyoda S, Chovatia M, Davies KM, Delmans M, Demura T, Dierschke T, Dolan L, Dorantes-Acosta AE, Eklund DM, Florent SN, Flores-Sandoval E, Fujiyama A, Fukuzawa H, Galik B, Grimanelli D, Grimwood J, Grossniklaus U, Hamada T, Haseloff J, Hetherington AJ, Higo A, Hirakawa Y, Hundley HN, Ikeda Y, Inoue K, Inoue SI, Ishida S, Jia Q, Kakita M, Kanazawa T, Kawai Y, Kawashima T, Kennedy M, Kinose K, Kinoshita T, Kohara Y, Koide E, Komatsu K, Kopischke S, Kubo M, Kyozuka J, Lagercrantz U, Lin SS, Lindquist E, Lipzen AM, Lu CW, De Luna E, Martienssen RA, Minamino N, Mizutani M, Mizutani M, Mochizuki N, Monte I, Mosher R, Nagasaki H, Nakagami H, Naramoto S, Nishitani K, Ohtani M, Okamoto T, Okumura M, Phillips J, Pollak B, Reinders A, Rovekamp M, Sano R, Sawa S, Schmid MW, Shirakawa M, Solano R, Spunde A, Suetsugu N, Sugano S, Sugiyama A, Sun R, Suzuki Y, Takenaka M, Takezawa D, Tomogane H, Tsuzuki M, Ueda T, Umeda M, Ward JM, Watanabe Y, Yazaki K, Yokoyama R, Yoshitake Y, Yotsui I, Zachgo S, Schmutz J (2017) Insights into land plant evolution garnered from the Marchantia polymorpha genome. Cell 171:287–304
Ohyama K, Fukuzawa H, Kohchi T, Shirai H, Sano T, Sano S, Umesono K, Shiki K, Takeuchi M, Chang Z, Aota S, Inokuchi H, Ozeki H (1986) Chloroplast gene organization deduced from complete sequence of liverwort, Marchantia polymorpha chloroplast DNA. Nature 322:572–574
Shinozaki K, Ohme M, Tanaka M, Wakasugi T, Hayashida N, Matsubayashi T, Zaita N, Chunwongse J, Obokata J, Yamaguchishinozaki K, Ohto C, Torazawa K, Meng BY, Sugita M, Deno H, Kamogashira T, Yamada K, Kusuda J, Takaiwa F, Kato A, Tohdoh N, Shimada H, Sugiura M (1986) The complete nucleotide-sequence of the tobacco chloroplast genome – its gene organization and expression. EMBO J 5:2043–2049
Chiyoda S, Ishizaki K, Kataoka H, Yamato KT, Kohchi T (2008) Direct transformation of the liverwort Marchantia polymorpha L. by particle bombardment using immature thalli developing from spores. Plant Cell Rep 27:1467–1473
Ishizaki K, Chiyoda S, Yamato KT, Kohchi T (2008) Agrobacterium-mediated transformation of the haploid liverwort Marchantia polymorpha L., an emerging model for plant biology. Plant Cell Physiol 49:1084–1091
Kubota A, Ishizaki K, Hosaka M, Kohchi T (2013) Efficient Agrobacterium-mediated transformation of the liverwort Marchantia polymorpha using regenerating thalli. Biosci Biotechnol Biochem 77:167–172
Tsuboyama S, Kodama Y (2014) AgarTrap: a simplified Agrobacterium-mediated transformation method for sporelings of the liverwort Marchantia polymorpha L. Plant Cell Physiol 55:229–236
Tsuboyama S, Nonaka S, Ezura H, Kodama Y (2018) Improved G-AgarTrap: a highly efficient transformation method for intact gemmalings of the liverwort Marchantia polymorpha. Sci Rep 8:10800
Ishizaki K, Nishihama R, Ueda M, Inoue K, Ishida S, Nishimura Y, Shikanai T, Kohchi T (2015) Development of gateway binary vector series with four different selection markers for the liverwort Marchantia polymorpha. PLoS One 10:e0138876
Mano S, Nishihama R, Ishida S, Hikino K, Kondo M, Nishimura M, Yamato KT, Kohchi T, Nakagawa T (2018) Novel gateway binary vectors for rapid tripartite DNA assembly and promoter analysis with various reporters and tags in the liverwort Marchantia polymorpha. PLoS One 13:e0204964
Ishizaki K, Johzuka-Hisatomi Y, Ishida S, Iida S, Kohchi T (2013) Homologous recombination-mediated gene targeting in the liverwort Marchantia polymorpha L. Sci Rep 3:1532
Nishihama R, Ishida S, Urawa H, Kamei Y, Kohchi T (2016) Conditional gene expression/deletion systems for Marchantia polymorpha using its own heat-shock promoter and Cre/loxP-mediated site-specific recombination. Plant Cell Physiol 57:271–280
Flores-Sandoval E, Dierschke T, Fisher TJ, Bowman JL (2016) Efficient and inducible use of artificial microRNAs in Marchantia polymorpha. Plant Cell Physiol 57:281–290
Sugano SS, Shirakawa M, Takagi J, Matsuda Y, Shimada T, Hara-Nishimura I, Kohchi T (2014) CRISPR/Cas9-mediated targeted mutagenesis in the liverwort Marchantia polymorpha L. Plant Cell Physiol 55:475–481
Sugano SS, Nishihama R, Shirakawa M, Takagi J, Matsuda Y, Ishida S, Shimada T, Hara-Nishimura I, Osakabe K, Kohchi T (2018) Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. PLoS One 13:e0205117
Tanaka D, Ishizaki K, Kohchi T, Yamato KT (2016) Cryopreservation of gemmae from the liverwort Marchantia polymorpha L. Plant Cell Physiol 57:300–306
Togawa T, Adachi T, Harada D, Mitani T, Tanaka D, Ishizaki K, Kohchi T, Yamato KT (2018) Cryopreservation of Marchantia polymorpha spermatozoa. J Plant Res 131:1047–1054
Chiyoda S, Linley PJ, Yamato KT, Fukuzawa H, Yokota A, Kohchi T (2007) Simple and efficient plastid transformation system for the liverwort Marchantia polymorpha L. suspension-culture cells. Transgenic Res 16:41–49
Maliga P (2004) Plastid transformation in higher plants. Annu Rev Plant Biol 55:289–313
Boehm CR, Ueda M, Nishimura Y, Shikanai T, Haseloff J (2016) A cyan fluorescent reporter expressed from the chloroplast genome of Marchantia polymorpha. Plant Cell Physiol 57:291–299
Chiyoda S, Kohchi T (2008) Plastid transformation system for suspension-culture cells and intact plants of liverwort Marchantia polymorpha L. Low Temp Sci 67:601–606. (in Japanese)
Acknowledgments
We thank Dr. F. Sato for the use of a biolistic delivery system. This work was supported in part by the grant “Knowledge Cluster Initiative” from MEXT.
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Chiyoda, S., Yamato, K.T., Kohchi, T. (2021). Plastid Transformation of Sporelings from the Liverwort Marchantia polymorpha L.. In: Maliga, P. (eds) Chloroplast Biotechnology. Methods in Molecular Biology, vol 2317. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1472-3_20
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DOI: https://doi.org/10.1007/978-1-0716-1472-3_20
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