Cytometric measurement of the DNA cell cycle in the presence of chlorophyll autofluorescence in marine eukaryotic phytoplankton by the blue-light excited dye YOYO-1
Frank J. Jochem1,*, Doris Meyerdierks2
1Institut für Meereskunde, Düsternbrooker Weg 20, D-24105 Kiel, Germany
2Universität Bremen, FB II Meeresbotanik, Postfach 330440, D-28334 Bremen, Germany
ABSTRACT: The novel blue-light excited fluorescent DNA dye YOYO-1® was tested on cultures of 8 eukaryotic phytoplankton species from the classes of Prymnesiophyceae, Bacillariophyceae, Coscinodiscophyceae, Prasinophyceae and
Dinophyceae, and the coccoid cyanobacterium Synechococcus sp. In all tested species, YOYO staining of formalin fixed cells allowed the differentiaton of cell cycle phases of G1, S, and G2+M in the presence of chlorophyll autofluorescence in
flow cytometric analyses. Diel cell cycle patterns of Emiliania huxleyi and Skeletonema costatum grown under a light:dark cycle and under continuous light could be established. Under the light/dark cycle, DNA replication
occurred during the night and cell division in the early morning in both species. However, S. costatum spent a longer fraction of their cell cycle in S and G2+M phases than E. huxleyi did. Changes in cellular chlorophyll
content and cell size could be observed in relation to cell cycle phasing. Cell division phasing was nearly lost under continuous light. Growth rates calculated from DNA cell cycle analyses agreed well with those established from cell counts.
KEY WORDS: Cell cycle · Cytometry · DNA · Growth rates