Skip to main content
Log in

Functional analysis of the eyespot in Chlamydomonas reinhardtii mutant ey 627, mt

  • Published:
Planta Aims and scope Submit manuscript

Abstract

The function of the eyespot in phototaxis of the flagellate green alga Chlamydomonas reinhardtii Dangeard was studied using quantitative reflection confocal laser scanning microscopy and photoelectric measurements. The reflective properties of the eyespot and the photoreceptor current of the C. reinhardtii eyespot mutant ey 627, mt were compared with those of Chlamydomonas strains possessing a well-developed eyespot. Under growth conditions in which strongly disorganized eyespots were observed in the mutant by electron microscopy, there was a significant reduction in the reflection intensity of the eyespot and in the amplitude ratio (500∶440 nm) of photoreceptor currents induced by flashes of 500- and 440-nm light in non-oriented cells. Photoelectrical responses of pre-oriented cells revealed that the latter effect is caused by an altered directional sensitivity of the antenna complex, whereas the functional state of the photoreceptor pigment is not strongly affected in mutant cells. Both the reflection intensity and the amplitude ratio of photoreceptor currents increased to the level of reference strains under conditions supporting the development of a well-organized eyespot in the mutant. Furthermore, incubation of the mutant with high concentrations of all-trans-retinal (10 μM), independent of whether carotenoid biosynthesis was inhibited or not, was found to increase the reflection intensity of the eyespot. An increase in the rate of photoorientation of the mutant occurred concomitant with the increase in the reflective properties of the mutant eyespot. These observations demonstrate the importance of an intact eyespot for interference reflection and absorption of phototactically active light, and thus for the directional sensitivity of the eyespot apparatus.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

HSM:

high-salt medium

Referencess

  • Beckmann, M., Hegemann, P. (1991) In vitro identification of rhodopsin in the green alga Chlamydomonas. Biochemistry 30, 3692–3697

    Google Scholar 

  • Boscov, J.S., Feinleib, M.E. (1979) Phototactic response of Chlamydomonas to flashes of light. II. Response of individual cells. Photochem. Photobiol. 30, 499–505

    Google Scholar 

  • Buder, J. (1917) Zur Kenntnis der phototaktischen Richtungsbewegung. Jahrb. Wiss. Bot. 58, 105–220

    Google Scholar 

  • Derguini, F., Mazur, P., Nakanishi, K., Starace, D.M., Saranak, J., Foster, K.W. (1991) All-trans-retinal is the chromophore bound to the photoreceptor of the alga Chlamydomonas reinhardtii. Photochem Photobiol. 54, 1017–1021

    Google Scholar 

  • Foster, K.W., Smyth, R.D. (1980) Light antennas in phototactic algae. Microbiol. Rev. 44, 572–630

    Google Scholar 

  • Foster, K.W., Saranak, J., Patel, N., Zarilli, G., Okabe, M., Kline, T., Nakanishi, K. (1984) A rhodopsin is the functional photoreceptor for phototaxis in the unicellular eukaryote Chlamydomonas. Nature 311, 756–759

    Google Scholar 

  • Foster, K.W., Saranak, J., Zarrilli, G. (1988) Autoregulation of rhodopsin synthesis in Chlamydomonas reinhardtii. Proc. Natl. Acad. Sci. USA 85, 6379–6383

    Google Scholar 

  • Harz, H., Hegemann, P. (1991) Rhodopsin-regulated calcium currents in Chlamydomonas. Nature 351, 489–491

    Google Scholar 

  • Hader, D.-P. (1979) Photomovement. In: Encyclopedia of plant physiology, N.S., vol. 7: Physiology of movements, pp. 268–309, Haupt, W., Feinleib, M.E., eds. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Hegemann, P., Hegemann, U., Foster, K.W. (1988) Reversible bleaching of Chlamydomonas reinhardtii rhodopsin in vivo. Photochem. Photobiol. 48, 123–128

    Google Scholar 

  • Hegemann, P., Gärtner, W., Uhl, R. (1991) All-trans retinal constitutes the functional chromophore in Chlamydomonas rhodopsin. Biophys. J. 60, 1477–1489

    Google Scholar 

  • Kreimer, G., Melkonian, M. (1990) Reflection confocal laser scanning microscopy of eyespots in flagellated green algae. Eur. J. Cell Biol. 53, 101–111

    Google Scholar 

  • Kreimer, G., Brohsonn, U., Melkonian, M. (1991a) Isolation and partial characterization of the photoreceptive organelle for phototaxis of a flagellate green alga. Eur. J. Cell Biol. 55, 318–327

    Google Scholar 

  • Kreimer, G., Kawai, H., Müller, D.G., Melkonian, M. (1991b) Reflective properties of the stigma in male gametes of Ectocarpus siliculosus (Phaeophyceae) studied by confocal laser scanning microscopy. J. Phycol. 27, 268–276

    Google Scholar 

  • Kreimer, G., Marner, F.-J., Brohsonn, U., Melkonian, M. (1991c) Identification of 11-cis and all-trans-retinal in the photoreceptive organelle of a flagellate green alga. FEBS-Lett. 293, 49–52

    Google Scholar 

  • Lien, T., Knutsen, G. (1976) Synchronized cultures of a cell wallless mutant of Chlamydomonas reinhardtii. Arch. Microbiol. 108, 189–194

    Google Scholar 

  • Lien, T., Knutsen, G. (1979) Synchronous growth of Chlamydomonas reinhardtii (Chlorophyceae): a review of optimal conditions. J. Phycol. 15, 191–200

    Google Scholar 

  • Litvin, F.F., Sineshchekov, O.A., Sineshchekov, V.A. (1978) Photoreceptor electric potential in the phototaxis of the alga Haematococcus pluvialis. Nature 271, 476–478

    Google Scholar 

  • Mast, S.O. (1927) Structure and function of the eye-spot in unicellular and colonial organisms. Arch. Protistenkd. 60, 197–220

    Google Scholar 

  • McFadden, G.I., Melkonian, M. (1986) Use of Hepes buffer for microalgal culture media and fixation for electron microscopy. Phycologia 25, 551–557

    Google Scholar 

  • Melkonian, M. (1975) The fine structure of zoospores of Fritschiella tuberosa lyeng (Chaetophorineae, Chlorophyceae) with special reference to the flagellar apparatus. Protoplasma 86, 391–404

    Google Scholar 

  • Melkonian, M. (1982) Effects of divalent cations on flagellar scales in the green flagellate Tetraselmis cordiformis. Protoplasma 111, 221–233

    Google Scholar 

  • Melkonian, M., Robenek, H. (1980) Eyespot membranes of Chlamydomonas reinhardtii: a freeze-fracture study. J. Ultrastruct. Res. 72, 90–102

    Google Scholar 

  • Melkonian, M., Robenek, H. (1984) The eyespot apparatus of flagellated green algae: a critical review. Prog. Phycol. Res. 3, 193–268

    Google Scholar 

  • Morel-Laurens, N.M.L., Bird, D.J. (1984) Effects of cell division on the stigma of wild-type and an “eyeless” mutant of Chlamydomonas. J. Ultrastruc. Res. 87, 46–61

    Google Scholar 

  • Morel-Laurens, N.M.L., Feinleib, M.E. (1983) Photomovement in an “eyeless” mutant of Chlamydomonas. Photochem. Photobiol. 37, 189–194

    Google Scholar 

  • Nultsch, W., Hader, D.-P. (1988) Photomovement in motile microorganisms-II. Photochem. Photobiol. 47, 837–869

    Google Scholar 

  • Nultsch, W., Throm, G., von Rimscha, I. (1971) Phototaktische Untersuchungen an Chlamydomonas reinhardtii Dangeard in homokontinuierlicher Kultur. Arch. Mikrobiol. 80, 351–369

    Google Scholar 

  • Pringsheim, E.G. (1963) Farblose Algen. Gustav Fischer Verlag, Stuttgart

    Google Scholar 

  • Reize, I.B., Melkonian, M. (1989) A new way to investigate living flagellated/ciliated cells in the light microscope: immobilization of cells in agarose. Bot. Acta 102, 145–151

    Google Scholar 

  • Rüffer, U., Nultsch, W. (1991) Flagellar photoresponses of Chlamydomonas cells held on micropipettes: IL Change in flagellar beat pattern. Cell Motil. Cytoskeleton 18, 269–278

    Google Scholar 

  • Sineshchekov, O.A. (1991a) Photoreception in unicellular flagellates: bioelectric phenomena in phototaxis. In: Light in biology and medicine, Proceedings of the III Conference of the European Society for Photobiology Budapest 1989, vol. 2, pp. 523–532, Douglas, R.H., Moan, J., Ronto, G., eds. Plenum Press, New York

    Google Scholar 

  • Sineshchekov, O.A. (1991b) Electrophysiology of photomovements in flagellated algae. In: Biophysics of photoreceptors and photomovements, pp. 191–202, Lenci, F. ed. Plenum Press, New York

    Google Scholar 

  • Sineshchekov, O.A., Govorunova, E.G., Litvin, F.F. (1989) Role of photosynthetic apparatus and stigma in the formation of spectral sensitivity of phototaxis in flagellated green algae. In Russ. Eng. abstr. Biofizika 34, 255–258

    Google Scholar 

  • Sineshchekov, O.A., Litvin, F.F., Keszthelyi, L. (1990) Two components of photoreceptor potential in phototaxis of the flagellated green alga Haematococcus pluvialis. Biophys. J. 57, 33–39

    Google Scholar 

  • Sineshchekov, O.A., Govorunova, E.G., Der, A., Keszthelyi, L., Nultsch, W. (1992) Photoelectric responses in phototactic flagellated algae measured in cell suspensions. J. Photochem. Photobiol. 13, 119–134

    Google Scholar 

  • Sueoka, N., Chiang, K.S., Kates, J.R. (1967) Deoxyribonucleic acid replication in meiosis of Chlamydomonas reinhardtii. I. Isotopic transfer experiments with a strain producing eight zoospores. J. Mol. Biol. 25, 47–66

    Google Scholar 

  • Surzycki, S. (1971) Synchronous grown cultures of Chlamydomonas reinhardtii. Methods Enzymol. 23, 67–73

    Google Scholar 

  • Takahashi, T., Yoshihara, K., Watanabe, M., Kubota, M., Johnson, R., Derguini, F., Nakanishi, K. (1991) Photoisomerization of retinal at 13-ene is important for phototaxis of Chlamydomonas reinhardtii: simultaneous measurements of phototactic and photophobic responses. Biochem. Biophys. Res. Commun. 178, 1273–1279

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

This study was supported by the Deutsche Forschungsgemeinschaft. O. A. Sineshchekov was supported by a Research Fellowship from the Alexander von Humboldt Foundation. The authors wish to thank U. Powalowski (Botanisches Institut, Universität zu Köln) for help with electron microscopy.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kreimer, G., Overländer, C., Sineshchekov, O.A. et al. Functional analysis of the eyespot in Chlamydomonas reinhardtii mutant ey 627, mt . Planta 188, 513–521 (1992). https://doi.org/10.1007/BF00197043

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00197043

Key words

Navigation