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A Simple Protocol for Thallus Culture-Based Genetic Transformation of the Liverwort Marchantia polymorpha

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Abstract

The liverwort Marchantia polymorpha, as the earliest land plant, is an important model system for studies of molecular evolution. Liverworts, which undergo a haplo-diplontic life cycle, spend most of their life as haploid gametophyte bodies, called thalli. In addition, the thalli can produce vegetative propagules, termed gemmae, conferring asexual reproduction in liverworts. Thus, to obtain sufficient isogenic tissues for genetic manipulation, the thallus is a suitable material. Here, we found that 23- to 25-d-old fully expanded thalli can be used for the genetic transformation with the pre-cultivation process, setting up the mature thalli pre-cultivation method. Next, we elucidated that 10- to 12-d-old young thalli can be used for the tissue-culture-based transformation in the absence of the pre-cultivation step, establishing the young thalli direct co-culture method without significant changes of transformation efficiency compared with the mature thalli pre-culture method. Overall, we set up the protocol using various stages of thalli of M. polymorpha with stable transformation efficiency. Mainly, we focused on reducing the hands-on labor time for performing the genetic transformation by widening the available stage of thalli for transformation or excluding the regenerating process of the thallus plantlets. In this report, we describe the detailed procedures employed under experimental conditions.

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The authors confirm that data supporting the findings in this study are available within the article and its supplementary materials.

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Acknowledgements

This work was supported by grants from the Basic Science Research Program (Project Number 2018R1A6A1A03025607) through the National Research Foundation (NRF) funded by the Ministry of Education, South Korea, and the Yonsei University Research Fund of 2020-22-0083 to WTK and the Young Scientist Grants Program (Project Number NRF-2020R1C1C1014389) and the Yonsei University Research Fund of 2020-12-0025 to DHS.

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Authors and Affiliations

  1. Department of Systems Biology and Division of Life Science, Yonsei University, Seoul, 03722, Korea

    Dong Hye Seo, Sojung Kim, Hyeon Ji Seo, Seung Eun Lee, Cheol Jin Lim, Hee Woong Yang, Li Hua Cui, Min Yong Ahn, Seong Gwan Yu, Na Hyun Cho, So Young Park, Ji Hee Lee & Woo Taek Kim

  2. Institute of Life Science and Biotechnology, Yonsei University, Seoul, 03722, Korea

    Dong Hye Seo, Sojung Kim, Hyeon Ji Seo, Seung Eun Lee, Cheol Jin Lim, Hee Woong Yang, Li Hua Cui, Min Yong Ahn, Seong Gwan Yu, Na Hyun Cho, So Young Park, Ji Hee Lee & Woo Taek Kim

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  1. Dong Hye Seo
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  2. Sojung Kim
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  3. Hyeon Ji Seo
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  5. Cheol Jin Lim
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  11. So Young Park
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Contributions

DHS, SK, HJS, SEL, CJL, HWY, LHC, MYA, SGY, NHC, SYP, and JHL: performed the experiments. DHS, SK, HJS, and WTK: analyzed the data. DHS: wrote the manuscript. WTK: supervised the project and complemented the writing.

Corresponding authors

Correspondence to Dong Hye Seo or Woo Taek Kim.

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Seo, D.H., Kim, S., Seo, H.J. et al. A Simple Protocol for Thallus Culture-Based Genetic Transformation of the Liverwort Marchantia polymorpha. J. Plant Biol. 65, 11–19 (2022). https://doi.org/10.1007/s12374-021-09339-w

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  • DOI: https://doi.org/10.1007/s12374-021-09339-w

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