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Optimized protoplast isolation and establishment of transient gene expression system for the Antarctic flowering plant Colobanthus quitensis (Kunth) Bartl.

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Abstract

Colobanthus quitensis is one of two terrestrial plants that grow in the maritime Antarctic. Despite its important ecological niche in extreme environments, the molecular mechanisms of its adaptation and tolerance have not been elucidated due to difficulties with genetic or transgenic approaches. For this reason, in many other plant species mesophyll protoplasts as a versatile cell-based system have been developed and used to analyze the biological functions of genes of interest. Here we report an optimized method of protoplast isolation from C. quitensis leaves. The main parameters evaluated to reach the highest protoplast yield were the use of a cell wall-degrading enzyme, an osmotic stabilizer, and different pH conditions. Moreover, transient expression and subcellular localization of proteins were validated by an immunoblot assay and a confocal microscopy, respectively, using C. quitensis protoplasts. Therefore, these results suggest that protoplasts can provide a useful cell-based system to facilitate the molecular, biochemical, and cellular characterizations of C. quitensis genes.

Key message

C. quitensis protoplasts can provide a physiologically relevant cell system to facilitate the molecular, biochemical, and cellular characterization of C. quitensis genes.

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Data availability

All sequence information of C. quitensis orthologous genes used in this study were submitted to GenBank under accession numbers MH0033823–MH0033828. All data are stored at Korea Polar Data Center (KPDC; http://kpdc.kopri.re.kr).

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Acknowledgements

This research was supported by a grant from the Korea Polar Research Institute (PE18290).

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Authors and Affiliations

  1. Department of Biotechnology, Duksung Women’s University, Seoul, 01369, Republic of Korea

    Ok-Kyoung Cha & Horim Lee

  2. Unit of Polar Genomics, Korea Polar Research Institute (KOPRI), Incheon, 21990, Republic of Korea

    Jungeun Lee & Hyoung Seok Lee

  3. Polar Science, University of Science & Technology, Daejeon, 34113, Republic of Korea

    Jungeun Lee & Hyoung Seok Lee

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  1. Ok-Kyoung Cha
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  2. Jungeun Lee
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  3. Hyoung Seok Lee
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  4. Horim Lee
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Contributions

JL and HL developed concept and supplied plant materials. JL, HSL, and HL designed the research. HL wrote the manuscript and OKC performed all experiments.

Corresponding author

Correspondence to Horim Lee.

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The authors declare no conflict of interest.

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Communicated by T. Winkelmann.

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Cha, OK., Lee, J., Lee, H.S. et al. Optimized protoplast isolation and establishment of transient gene expression system for the Antarctic flowering plant Colobanthus quitensis (Kunth) Bartl.. Plant Cell Tiss Organ Cult 138, 603–607 (2019). https://doi.org/10.1007/s11240-019-01651-1

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