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Open AccessArticle

Endosymbiotic Green Algae in Paramecium bursaria: A New Isolation Method and a Simple Diagnostic PCR Approach for the Identification

1
Research Department for Limnology Mondsee, Leopold-Franzens-University of Innsbruck, A-5310 Mondsee, Austria
2
Albrecht-von-Haller-Institute of Plant Sciences, Experimental Phycology and Culture Collection of Algae, Georg-August-University of Göttingen, D-37073 Göttingen, Germany
3
Applied Ecology and Phycology, University of Rostock, D-18059 Rostock, Germany
*
Author to whom correspondence should be addressed.
Diversity 2020, 12(6), 240; https://doi.org/10.3390/d12060240
Received: 15 May 2020 / Revised: 6 June 2020 / Accepted: 10 June 2020 / Published: 12 June 2020
(This article belongs to the Special Issue Biodiversity of Ciliates and their Symbionts)
Paramecium bursaria is a single-celled model organism for studying endosymbiosis among ciliates and green algae. Most strains of P. bursaria bear either Chlorella variabilis or Micractinium conductrix as endosymbionts. Both algal genera are unicellular green algae characterized by cup-shaped chloroplasts containing a single pyrenoid and reproduction by autospores. Due to their size and only few morphological characteristics, these green algae are very difficult to discriminate by microscopy only. Their cultivation is laborious and often unsuccessful, but we developed a three-step isolation method, which provided axenic cultures of endosymbionts. In addition to the time-consuming isolation, we developed a simple diagnostic PCR identification method using specific primers for C. variabilis and M. conductrix that provided reliable results. One advantage of this approach was that the algae do not have to be isolated from their host. For a comparative study, we investigated 19 strains of P. bursaria from all over the world (new isolates and available laboratory strains) belonging to the five known syngens (R1–R5). Six European ciliate strains belonging to syngens R1 and R2 bore M. conductrix as endosymbiont whereas C. variabilis was discovered in syngens R1–R5 having worldwide origins. Our results reveal the first evidence of C. variabilis as endosymbiont in P. bursaria in Europe. View Full-Text
Keywords: Chlorella variabilis; Micractinium conductrix; diagnostic PCR; Paramecium; ciliate–algae symbiosis Chlorella variabilis; Micractinium conductrix; diagnostic PCR; Paramecium; ciliate–algae symbiosis
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MDPI and ACS Style

Spanner, C.; Darienko, T.; Biehler, T.; Sonntag, B.; Pröschold, T. Endosymbiotic Green Algae in Paramecium bursaria: A New Isolation Method and a Simple Diagnostic PCR Approach for the Identification. Diversity 2020, 12, 240.

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