Antifreeze Water-Rich Dormant Cysts of the Terrestrial Ciliate Colpoda cucullus Nag-1 at −65 ℃: Possible Involvement of Ultra-Antifreeze Polysaccharides

Tatsuomi Matsuoka; Yoichiro Sogame; Rikiya Nakamura; Yuya Hasegawa; Mikihiko Arikawa; Futoshi Suizu

Antifreeze Water-Rich Dormant Cysts of the Terrestrial Ciliate Colpoda cucullus Nag-1 at −65 ℃: Possible Involvement of Ultra-Antifreeze Polysaccharides

Publication date: 2021

Acta Protozoologica, 2020, Volume 59, Issue 3-4, pp. 141 - 147

https://doi.org/10.4467/16890027AP.20.011.13266

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Antifreeze Water-Rich Dormant Cysts of the Terrestrial Ciliate Colpoda cucullus Nag-1 at −65 ℃: Possible Involvement of Ultra-Antifreeze Polysaccharides

Abstract

We found that the water-rich (osmolality below 0.052 Osm/l) wet resting cysts of the soil ciliate Colpoda cucullus Nag-1 were tolerant to extremely low temperature (−65℃). When cell fluid obtained from the resting cysts was cooled at −65℃, small particles of ice crystals did not grow into large ice crystals. At −65℃, the cysts shrank due to an outflow of water, because a vapor pressure difference was produced between the cell interior and freezing surrounding medium. The osmolality of these shrunk cells was estimated 0.55 Osm/l, and the freezing point depression of the shrunk cell fluid was estimated to be 1.02℃. Hence, the antifreeze ability of wet cysts at −65℃can not be explained by freezing point depression due to elevation of cytoplasmic osmolality.

The cytoplasm of resting cysts was vividly stained red with periodic acid-Schiff (PAS) and stained purple with toluidine blue. On the other hand, the excystment-induced cysts were not stained with PAS, and exhibited a loss of the antifreeze activity. PAS staining of SDSPAGE gel obtained from encysting Colpoda cells showed that a large amount of PAS-positive macromolecules accumulated as the encystment stage progressed. These results suggest that antifreeze polysaccharides may be involved in the antifreeze activity of C. cucullus Nag-1 dormant forms.

Keywords

water-rich resting cyst, antifreeze, polysaccharides, osmolarity

References

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Information

Information: Acta Protozoologica, 2020, Volume 59, Issue 3-4, pp. 141 - 147

DOI: https://doi.org/10.4467/16890027AP.20.011.13266

Article type: Original article

Authors

Tatsuomi Matsuoka

Department of Biological Science, Faculty of Science, Kochi University, Kochi, Japan

Yoichiro Sogame

National Institute of Technology Fukushima College, Iwaki, Fukushima Japan

Rikiya Nakamura

Department of Biological Science, Faculty of Science, Kochi University, Kochi, Japan

Yuya Hasegawa

Department of Biological Science, Faculty of Science, Kochi University, Kochi, Japan

Mikihiko Arikawa

Department of Biological Science, Faculty of Science, Kochi University, Kochi, Japan

Futoshi Suizu

Oncology Pathology, Department of Pathology and Host-Defense, Faculty of Medicine, Kagawa University, Takamatsu 761-0793, Japan

Published at: 2021

Received at: 29.09.2020

Accepted at: 02.12.2020

Article status: Open

Licence: CC BY-NC-ND

Percentage share of authors:

Tatsuomi Matsuoka (Author) - 16%

Yoichiro Sogame (Author) - 16%

Rikiya Nakamura (Author) - 16%

Yuya Hasegawa (Author) - 16%

Mikihiko Arikawa (Author) - 16%

Futoshi Suizu (Author) - 20%

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English

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<p> Antifreeze Water-Rich Dormant Cysts of the Terrestrial Ciliate <em>Colpoda cucullus </em>Nag-1 at −65 ℃: Possible Involvement of Ultra-Antifreeze Polysaccharides</p>

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