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Development of desiccation tolerance and vitrification by preculture treatment in suspension-cultured cells of the liverwort Marchantia polymorpha

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Abstract

Some cultured plant cells are able to acquire tolerance to various stresses when they are cultured under suitably controlled conditions. Induction of a high level of desiccation tolerance in suspension-cultured cells of the liverwort Marchantia polymorpha was examined for studying the mechanisms of desiccation tolerance and vitrification at the cellular level. Desiccation tolerance level of cells was very low and the survival rate was less than 10% after exposure to drying below 0.1 g H2O g−1 dry weight (DW). Preculture treatment in 0.5 M sucrose medium was the most effective method for inducing a high level of desiccation tolerance in cells and the survival rate was 87% even after being desiccated to below 0.1 g H2O g−1 DW. Preculture treatment caused alteration of cell structures and accumulation of a large amount of sucrose and newly synthesized proteins in cells. Abundant sucrose and preculture-induced proteins were necessary for full development of desiccation tolerance in the cells. When water content decreased to below 0.1 g H2O g−1 DW, desiccation-tolerant cells that had been precultured were vitrified above 0°C and maintained stable viability. We have succeeded in the induction of desiccation tolerance that allows formation of intracellular glass with cell viability at ambient temperatures by controlling culture conditions, and our results suggest that suspension-cultured cells of M. polymorpha are useful for studying cellular mechanisms for the development of desiccation tolerance and the stabilization of vitrified cells.

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Abbreviations

ABA:

Abscisic acid

DSC:

Differential scanning calorimetry

LEA:

Late embryogenesis abundant

T g :

Glass transition temperature

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Acknowledgments

This study was partly supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (Grant no. 18658040, Y. S.). We thank Dr. Yasuko Kaneko for her support with the electron microscopy.

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  1. Graduate School of Science and Engineering, Department of Regulation-Biology, Faculty of Science, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama City, Saitama, 338-8570, Japan

    Rie Hatanaka & Yasutake Sugawara

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  1. Rie Hatanaka
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  2. Yasutake Sugawara
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Correspondence to Yasutake Sugawara.

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Hatanaka, R., Sugawara, Y. Development of desiccation tolerance and vitrification by preculture treatment in suspension-cultured cells of the liverwort Marchantia polymorpha . Planta 231, 965–976 (2010). https://doi.org/10.1007/s00425-010-1101-5

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