Assessing phagotrophy in the mixotrophic ciliate Paramecium bursaria using GFP-expressing yeast cells

Abstract We used cells of the yeast Saccharomyces cerevisiae expressing green fluorescent protein (GFP) as fluorescently labelled prey to assess the phagocytic activities of the mixotrophic ciliate Paramecium bursaria, which harbours symbiotic Chlorella-like algae. Because of different fluorescence...

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Bibliographic Details
Published in:FEMS microbiology letters 2017-07, Vol.364 (12)
Main Authors: Miura, Takashi, Moriya, Hisao, Iwai, Sosuke
Format: Article
Language:English
Subjects:
Algae
Baking yeast
Chlorella vulgaris - physiology
Chlorophyll
Ciliates
Digestion
Endosymbionts
Fluorescence
Fluorescence microscopy
Food vacuoles
Green fluorescent protein
Green Fluorescent Proteins - genetics
Ingestion
Microbiology
Paramecium - genetics
Paramecium - growth & development
Paramecium - physiology
Paramecium bursaria
Phagocytes
Phagocytosis
Photosynthesis
Prey
Protein expression
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Symbiosis
Vacuoles
Yeast
Yeasts
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Summary:Abstract We used cells of the yeast Saccharomyces cerevisiae expressing green fluorescent protein (GFP) as fluorescently labelled prey to assess the phagocytic activities of the mixotrophic ciliate Paramecium bursaria, which harbours symbiotic Chlorella-like algae. Because of different fluorescence spectra of GFP and algal chlorophyll, ingested GFP-expressing yeast cells can be distinguished from endosymbiotic algal cells and directly counted in individual P. bursaria cells using fluorescence microscopy. By using GFP-expressing yeast cells, we found that P. bursaria altered ingestion activities under different physiological conditions, such as different growth phases or the presence/absence of endosymbionts. Use of GFP-expressing yeast cells allowed us to estimate the digestion rates of live prey of the ciliate. In contrast to the ingestion activities, the digestion rate within food vacuoles was not affected by the presence of endosymbionts, consistent with previous findings that food and perialgal vacuoles are spatially and functionally separated in P. bursaria. Thus, GFP-expressing yeast may provide a valuable tool to assess both ingestion and digestion activities of ciliates that feed on eukaryotic organisms. Phagotrophy of the mixotrophic ciliate Paramecium bursaria harbouring symbiotic algae was monitored using yeast cells expressing green fluorescent protein.
ISSN:1574-6968
0378-1097
1574-6968
DOI:10.1093/femsle/fnx117