, Volume 201, Issue 1–2, pp 45–52 | Cite as

Discovery of signaling effect of UV-B/C light in the extended UV-A/blue-type action spectra for step-down and step-up photophobic responses in the unicellular flagellate algaEuglena gracilis

  • S. Matsunaga
  • T. Hori
  • T. Takahashi
  • M. Kubota
  • M. Watanabe
  • K. Okamoto
  • K. Masuda
  • M. Sugai


Cultures of unicellular algal flagellateEuglena gracilis grown in different conditions were subjected to action spectroscopy for step-down and step-up photophobic responses, respectively. The spectral region was extended into the UV-B/C as well as in the UV-A and visible regions with the Okazaki Large Spectrograph as the monochromatic light source. The photophobic responses of the cells were measured with an individual-cell assay method with the aid of a computerized video motion analyzer. In the UV-A and visible regions, the shapes of the action spectra were the so-called UV-A/blue type. In the newly studied UV-B/C region, new action peaks were found at 270 nm for the step-down response and at 280 nm for the step-up one. The absorption spectrum of flavin adenine dinucleotide (FAD) appeared to fit the action spectrum for the step-up response, whereas the shape of the step-down action spectrum, which has a UV-A peak (at 370 nm) higher than the blue peak (at 450 nm), appeared to be mimicked by the absorption spectrum of a mixed solution of 6-biopterin and FAD. These observations might also account for the fact that the UV-B/C peak wavelength at 270 nm of the action spectrum for the step-down response is shorter by 10 nm than the action spectrum for the step-up response at 280 nm.


Action spectra Blue light Euglena Photophobic responses Ultraviolet light 



flavin adenine dinucleotide


spectral full width at half maximum


National Institute for Basic Biology


Okazaki Large Spectrograph


paraflagellar body


ultraviolet light of spectral region between 320 and 400 nm


ultraviolet light of spectral region between 190 and 320 nm


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ahmad M, Cashmore AR (1993)HY4 gene ofA. thaliana encodes a protein with characteristics of a blue-light photoreceptor. Nature 336: 162–166Google Scholar
  2. Barghigiani C, Colombetti G, Franchini B, Lend F (1979) Photobehavior ofEuglena gracilis: action spectrum for the step-down photophobic response of individual cells. Photochem Photobiol 29: 1015–1019Google Scholar
  3. Baskin TI, Iino M (1987) An action spectrum in the blue and ultraviolet for phototropism inAlfalfa. Photochem Photobiol 46: 127–136Google Scholar
  4. Benedetti PA, Lenci F (1977) In vivo microspectrofluorometry of photoreceptor pigments inEuglena gracilis. Photochem Photobiol 26: 315–318Google Scholar
  5. Brodhun B, Häder DP (1990) Photoreceptor proteins and pigments in the paraflagellar body of the flagellateEuglena gracilis. Photochem Photobiol 52: 865–871Google Scholar
  6. Checcucci A, Colombetti G, Ferrara R, Lenci F (1976) Action spectra for photoaccumulation of green and colorlessEuglena: evidence for identification of receptor pigments. Photochem Photobiol 23: 51–54PubMedGoogle Scholar
  7. Colombetti G, Häder DP, Lenci F, Quaglia M (1982a) Phototaxis inEuglena gracilis. Curr Microbiol 4: 281–284Google Scholar
  8. —, Lenci F, Diehn B (1982b) Responses to photic, chemical and mechanical stimuli. In: Buetow DE (ed) The biology ofEuglena, vol 3. Academic Press, New York, pp 169–195Google Scholar
  9. Deininger W, Kröger P, Hegemann U, Lottspeich F, Hegemann P (1995) Chlamyrhodopsin represents a new type of sensory photoreceptor. EMBO J 14: 5849–5858PubMedGoogle Scholar
  10. Diehn B (1969a) Action spectra of the phototactic responses inEuglena. Biochim Biophys Acta 177: 136–143PubMedGoogle Scholar
  11. — (1969b) Phototactic response ofEuglena to single and repetitive pulses of actinic light: orientation time and mechanism. Exp Cell Res 56: 375–381PubMedGoogle Scholar
  12. — (1972) The receptor/effector system of phototaxis inEuglena. Acta Protozool 11: 325–332Google Scholar
  13. Erata M, Kubota M, Takahashi T, Inoue I, Watanabe M (1995) Ultrastructure and phototactic action spectra of two genera of cryptophyte flagellate algaeCryptomonas andChroomonas. Protoplasma 188: 258–266Google Scholar
  14. Foster KW, Saranak J, Patel N, Zarilli G, Okabe M, Kline T, Nakanishi K (1984) A rhodopsin is the functional photoreceptor for phototaxis in the unicellular eukaryoteChlamydotnonas. Nature 311: 756–759PubMedGoogle Scholar
  15. Galland P, Keiner P, Dörnemann D, Senger H, Brodhun B, Häder DP (1990) Pterin- and flavin-like fluorescence associated with isolated flagella ofEuglena gracilis. Photochem Photobiol 51: 675–680Google Scholar
  16. -, Geiß D, Sineshchekov VA, Hohl N, Senger H (1994) UV-blue-light-reception inPhycomyces andEuglena: a role for pterins and flavins. Photochem Photobiol Suppl 59: 69SGoogle Scholar
  17. Geiss D, Senger H, Galland P (1997) Pigments associated with the flagellum ofEuglena gracilis. J Plant Res 110: 393–403Google Scholar
  18. Ghetti F, Colombetti G, Lenci F, Campani E, Polacco E, Quaglia M (1985) Fluorescence ofEuglena gracilis photoreceptor pigment: an in vivo microfluorometric study. Photochem Photobiol 42: 29–33Google Scholar
  19. Gualtieri P (1993)Euglena gracilis: is the photoreception enigma solved? J Photochem Photobiol B 19: 3–14Google Scholar
  20. Häder DP, Reinecke E (1991) Phototactic and polarotactic responses of the photosynthetic flagellate,Euglena gracilis. Acta Protozool 30: 13–18Google Scholar
  21. Halldal P (1961) Ultraviolet action spectra of positive and negative phototaxis inPlatymonas subcordiformis. Physiol Plant 14: 133–139Google Scholar
  22. Hand WG, Davenport D (1970) The experimental analysis of phototaxis and photokinesis in flagellates. In: Halldal P (ed) Photobiology of microorganisms. Wiley-Interscience, London, pp 253–282Google Scholar
  23. Hashimoto T, Yatsuhashi H, Kato H (1982) A high-sensitivity photon density meter for monochromatic light. In: Abstracts of the Annual Meeting of the Japanese Society Plant Physiology, p 38Google Scholar
  24. Inoue Y, Watanabe M (1984) Perithecial formation inGerasinospora reticulispora VII: action spectra in UV region for the photoinhibition of photoinductive effect brought by blue light. Plant Cell Physiol 25: 107–117Google Scholar
  25. Kawai H, Kubota M, Kondo T, Watanabe M (1991) Action spectra for phototaxis in zoospores of the brown algaPseudochorda gracilis. Protoplasma 161: 17–22Google Scholar
  26. Kondo T, Kubota M, Aono Y, Watanabe M (1988) A computerized video system to automatically analyze movements of individual cell and its application to the study of circadian rhythms in phototaxis and motility inChlamydomonas reinhardtii. Protoplasma Suppl 1: 185–192Google Scholar
  27. Kreimer G (1994) Cell biology of phototaxis in flagellate algae. Int Rev Cytol 148: 229–310Google Scholar
  28. Kroger P, Hegemann P (1994) Photophobic responses and phototaxis inChlamydomonas are triggered by a single rhodopsin photoreceptor. FEBS Lett 341: 5–9PubMedGoogle Scholar
  29. Kuznicki L, Walne PL (1989) The relationship between step-up and step-down photophobic responses inEuglena gracilis. Acta Protozool 28: 215–229Google Scholar
  30. Lawson MA, Zacks DN, Derguini F, Nakanishi K, Spudich JL (1991) Retinal analog restoration of photophobic responses in a blindChlamydomonas reinhardtii mutant. Biophys J 60: 1490–1498PubMedGoogle Scholar
  31. Lin C, Robertson DE, Ahmad M, Raibekas AA, Jorns MS, Dutton PL, Cashmore AR (1995) Association of flavin adenine dinucreotide with theArabidopsis blue light receptor CRY1. Science 269: 968–969PubMedGoogle Scholar
  32. Lipson ED (1995) Action spectroscopy: methodology. In: Horspool W, Song PS (eds) Handbook of organic photochemistry and photobiology. CRC Press, Boca Raton, pp 1257–1275Google Scholar
  33. Mikolajczyk E (1984) Photophobic responses inEuglena 1: effects of excitation wavelength and external medium on the step-up response of light- and dark-grownEuglena gracilis. Acta Protozool 23: 1–10Google Scholar
  34. —, Diehn B (1975) The effect of potassium iodide on photophobic responses inEuglena: evidence for two photoreceptor pigments. Photochem Photobiol 22: 269–271PubMedGoogle Scholar
  35. — — (1976) Light-induced body movement ofEuglena gracilis coupled to flagellar photophobic response by mechanical stimulation. J Protozool 23: 144–147Google Scholar
  36. — — (1978) Morphological alternations inEuglena gracilis induced by treatment with CTAB (cetyltrimethylammonium bromide) and Triton X-100: correlations with effects on photophobic behavioral responses. J Protozool 25: 461–470Google Scholar
  37. Nultsch W, Throm G, Rimscha I (1971) Phototaktische Untersuchungen anChlamydomonas reinhardii Dangeard in homokontinuierlicher Kultur. Arch Mikrobiol 80: 351–369Google Scholar
  38. Schäfer E, Fukshansky L (1984) Action spectroscopy. In: Smith H, Holmes MG (eds) Techniques in photomorphogenesis. Academic Press, London, pp 109–129Google Scholar
  39. Schmidt W, Galland P, Senger H, Furuya M (1990) Microspectrophotometry ofEuglena gracilis: pterin- and flavin-like fluorescence in the paraflagellar body. Planta 182: 375–381Google Scholar
  40. Shimmen T (1981) Quantitative studies on step-down photophobic response ofEuglena in an individual cell. Protoplasma 106: 37–48Google Scholar
  41. Sineshchekov VA, Geiss D, Sineshchekov OA, Galland P, Senger H (1994) Fluorometric characterization of pigments associated with isolated flagella ofEuglena gracilis: evidence for energy migration. J Photochem Photobiol B 23: 225–237Google Scholar
  42. Sugai M, Furuya M (1985) Action spectrum in ultraviolet and blue light region for the inhibition of red-light-induced spore germination inAdiantum capillus-veneris L. Plant Cell Physiol 26: 953–956Google Scholar
  43. —, Tomizawa K, Watanabe M, Furuya M (1984) Action spectrum between 250 and 800 nanometers for the photoinduced inhibition of spore germination inPteris vittata. Plant Cell Physiol 25: 205–212Google Scholar
  44. Suzaki T, Williamson RE (1983) Photoresponse of a colorless euglenoid flagellate,Astasia longa. Plant Sci Lett 32: 101–107Google Scholar
  45. Takahashi T (1991) Automated measurement of movement responses in Halobacteria. In: Häder DP (ed) Image analysis in biology. CRC Press, Boca Raton, pp 315Google Scholar
  46. —, Kobatake Y (1982) Computer-linked automated method for measurement of the reversal frequency in phototaxis ofHalobacterium halobium. Cell Struct Funct 7: 183–192Google Scholar
  47. —, Yoshihara K, Watanabe M, Kubota M, Johnson R, Derguini F, Nakanishi K (1991) Photoisomerization of retinal at 13-ene is important for phototaxis ofChlamydomonas reinhardtii: simultaneous measurements of phototactic and photophobic responses. Biochem Biophys Res Commun 178: 1273–1279PubMedGoogle Scholar
  48. Ueda T, Mori Y, Nakagaki T, Kobatake Y (1988) Action spectra for superoxide generation and UV and visible light photoavoidance in plasmodia ofPhysarum polycephalum. Photochem Photobiol 48: 705–709Google Scholar
  49. Watanabe M (1991) High-fluence rate monochromatic light sources, computerized analysis of cell movements, and microbeam irradiation of a moving cell: current experimental methodology at the Okazaki Large Spectrograph. In: Lenci F, Ghetti F, Colombetti G, Häder DP, Song PS (eds) Biophysics of photoreceptors and photomovements in microorganisms. Plenum, New York pp 327–337Google Scholar
  50. — (1995) Action spectroscopy: photomovement and photomorphogenesis spectra. In: Horspool W, Song PS (eds) Handbook of organic photochemistry and photobiology. CRC Press, Boca Raton, pp 1276–1288Google Scholar
  51. —, Furuya M, Miyoshi Y, Inoue Y, Iwahashi I, Matsumoto K (1982) Design and performance of the Okazaki Large Spectrograph for photobiological research. Photochem Photobiol 36: 491–498Google Scholar

Copyright information

© Springer-Verlag 1998

Authors and Affiliations

  • S. Matsunaga
    • 1
  • T. Hori
    • 1
  • T. Takahashi
    • 2
  • M. Kubota
    • 3
  • M. Watanabe
    • 3
  • K. Okamoto
    • 4
  • K. Masuda
    • 4
  • M. Sugai
    • 4
  1. 1.Institute of Biological SciencesUniversity of TsukubaIbarakiJapan
  2. 2.Japan Advanced Institute of Science and TechnologyIshikawa
  3. 3.National Institute for Basic BiologyAichi
  4. 4.Department of Biology, Faculty of ScienceToyama UniversityToyama

Personalised recommendations