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Plasmolysis effects and osmotic potential of two phylogenetically distinct alpine strains of Klebsormidium (Streptophyta)

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Abstract

The osmotic potential and effects of plasmolysis were investigated in two different Klebsormidium strains from alpine habitats by incubation in 300–2,000 (3,000) mM sorbitol. Several members of this genus were previously found to tolerate desiccation in the vegetative state yet information was lacking on the osmotic potentials of these algae. The strains were morphologically determined as Klebsormidium crenulatum and Klebsormidium nitens. These species belong to distinct clades, as verified by phylogenetic analysis of the rbcL gene. K. crenulatum is part of to the K. crenulatum/mucosum (‘F’ clade) and K. nitens of the ‘E2’ clade. Plasmolysis occurred in K. crenulatum at 800 mM sorbitol (961 mOsmol kg−1, Ψ = −2.09 MPa) and in K. nitens at 600 mM sorbitol (720 mOsmol kg−1, Ψ = −1.67 MPa). These are extraordinarily high osmotic values (very negative osmotic potentials) compared with values reported for other green algae. In K. crenulatum, the maximum photosynthetic rate (Pmax) in the light-saturated range was 116 μmol O2 h−1 mg−1 chl a. Incubation in 1,000 mM sorbitol decreased Pmax to 74.1% of the initial value, whereas 2,000 mM sorbitol (Ψ = −5.87 MPa) lead to an almost complete loss of oxygen production. In K. nitens, Pmax was 91 μmol O2 h−1 mg−1 chl a under control conditions and incubation in 800 mM sorbitol did not decrease Pmax, 2,000 mM sorbitol decreased Pmax only to about 62.6% of the initial value whereas 3,000 mM sorbitol stopped oxygen evolution. This indicated a broader amplitude for photosynthesis in the examined strain of K. nitens. Control samples and samples plasmolysed for 3 h in 800 mM sorbitol (K. nitens), 1,000 mM sorbitol (K. crenulatum), or 2,000 mM sorbitol were investigated by transmission electron microscopy after chemical or high-pressure freeze fixation. In cells undergoing plasmolysis the protoplasts were retracted from the cell wall, the cytoplasm appeared dense, vacuoles were small and fragmented, and the cytoplasm was filled with ribosomes. Thin cytoplasmic strands were connected to the cell wall; 2,000 mM sorbitol increased the effect. The content of soluble carbohydrates in these two strains was investigated by HPLC, as this is one known mechanism for cells to maintain high osmotic pressure of the cytosol. Both Klebsormidium species contained diverse soluble carbohydrates, including a dominant mixed peak of unidentified oligosaccharides, and more minor amounts of raffinose, sucrose, glucose, xylose, galactose, mannose, inositol, fructose, glycerol, mannitol, and sorbitol. The total content of soluble carbohydrates was approximately 1.2% of the dry weight, indicating that this is not a major factor contributing to the high osmotic potential in these strains of Klebsormidium.

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Acknowledgments

Prof. U. Lütz-Meindl, University of Salzburg, Austria is kindly acknowledged for providing access to her Leica high-pressure freezing device. We thank Mag. A. Andosch, University of Salzburg for technical help with the freeze substitution. We would like to thank Prof. U. Karsten, University of Rostock, Germany for isolating the Klebsormidium strain from Innsbruck, for critically reading the manuscript and several helpful discussions. Dr. M. Roleda, AWI Bremerhaven, Germany is acknowledged for help in calculating the P–E curve parameters. The rbcL data were generated during a scientific stay of A.H. in L.A.L.’s laboratory, supported by the Office of International Affairs of the University of Innsbruck and a US NASA Exobiology NNX08AX20G grant to Z. Cardon, L. Lewis, and H. Frank. The study has been supported by a Tyrolean Science Fund Project AP 717029 to A.H.

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  1. Institute of Botany, Functional Plant Biology, University of Innsbruck, Sternwartestrasse 15, 6020, Innsbruck, Austria

    Franziska Kaplan, Johann Wastian & Andreas Holzinger

  2. Department of Ecology and Evolutionary Biology, University of Connecticut, Storrs, CT, 06269-3043, USA

    Louise A. Lewis

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Kaplan, F., Lewis, L.A., Wastian, J. et al. Plasmolysis effects and osmotic potential of two phylogenetically distinct alpine strains of Klebsormidium (Streptophyta). Protoplasma 249, 789–804 (2012). https://doi.org/10.1007/s00709-011-0324-z

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