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Protoplasma

, Volume 250, Issue 1, pp 261–272 | Cite as

Ultrastructural analysis of flagellar development in plurilocular sporangia of Ectocarpus siliculosus (Phaeophyceae)

  • Gang Fu
  • Chikako Nagasato
  • Toshiaki Ito
  • Dieter G. Müller
  • Taizo MotomuraEmail author
Original Article

Abstract

Flagellar development in the plurilocular zoidangia of sporophytes of the brown alga Ectocarpus siliculosus was analyzed in detail using transmission electron microscopy and electron tomography. A series of cell divisions in the plurilocular zoidangia produced the spore-mother cells. In these cells, the centrioles differentiated into flagellar basal bodies with basal plates at their distal ends and attached to the plasma membrane. The plasma membrane formed a depression (flagellar pocket) into where the flagella elongated and in which variously sized vesicles and cytoplasmic fragments accumulated. The anterior and posterior flagella started elongating simultaneously, and the vesicles and cytoplasmic fragments in the flagellar pocket fused to the flagellar membranes. The two flagella (anterior and posterior) could be clearly distinguished from each other at the initial stage of their development by differences in length, diameter and the appendage flagellar rootlets. Flagella continued to elongate in the flagellar pocket and maintained their mutually parallel arrangement as the flagellar pocket gradually changed position. In mature zoids, the basal part of the posterior flagellum (paraflagellar body) characteristically became swollen and faced the eyespot region. Electron dense materials accumulated between the axoneme and the flagellar membrane, and crystallized materials could also be observed in the swollen region. Before liberation of the zoospores from the plurilocular zoidangia, mastigoneme attachment was restricted to the distal region of the anterior flagellum. Structures just below the flagellar membrane that connected to the mastigonemes were clearly visible by electron tomography.

Keywords

Brown algae Ectocarpus Flagellar differentiation Transmission electron microscopy Electron tomography 

Abbreviations

TEM

Transmission electron microscope

ET

Electron tomography

MT

Microtubule

EDM

Electron-dense materials

CM

Crystallized materials

Notes

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

709_2012_405_Fig10_ESM.jpg (40 kb)
Supplementary Fig. 1

a, b Vesicles in the flagellar lumen (arrows). Arrowheads indicate the flagellar membrane (JPEG 39 kb)

709_2012_405_MOESM1_ESM.tif (1.4 mb)
High resolution (TIFF 1441 kb)
Video 1

Paraflagellar body of the posterior flagellum. The video is composed of 60 adjacent slices (each 1.2-nm thickness) from a dual-axis tomogram corresponding to a paraflagellar body. Regular arrangement of crystalized materials can be detected. Scale bars = 50 nm. (MP4 220 kb)

References

  1. Andersen RA (2004) Biology and systematics of heterokont and haptophyte algae. Am J Bot 91:1508–1522PubMedCrossRefGoogle Scholar
  2. Baker JRJ, Evans LV (1973a) The ship-fouling alga Ectocarpus. I. Ultrastructure and cytochemistry of plurilocular reproductive stages. Protoplasma 77:1–13CrossRefGoogle Scholar
  3. Baker JRJ, Evans LV (1973b) The ship-fouling alga Ectocarpus. II. Ultrastructure of the unilocular reproductive stages. Protoplasma 77:181–189CrossRefGoogle Scholar
  4. Beech PL, Heimann K, Melkonian M (1991) Development of the flagellar apparatus during the cell cycle in unicellular algae. Protoplasma 164:23–37CrossRefGoogle Scholar
  5. Bouck GB (1969) Extracellular microtubules. The origin, structure, and attachment of flagellar hairs in Fucus and Ascophyllum antherozoids. J Cell Biol 40:446–460PubMedCrossRefGoogle Scholar
  6. Bui KH, Pigino G, Ishikawa T (2011) Three-dimensional structural analysis of eukaryotic flagella/cilia by electron cryo-tomography. J Synchrotron Radiat 18:2–5PubMedCrossRefGoogle Scholar
  7. Chang P, Stearns T (2000) δ-tubulin and ε-tubulin: two new human centrosomal tubulins reveal new aspects of centrosome structure and function. Nat Cell Biol 2:30–35PubMedCrossRefGoogle Scholar
  8. Clayton MN (1989) Brown algae and chromophyte phylogeny. In: Green JC, Leadbeater BSC and Diver WL (eds) The chromophyte alge: problems and perspectives. Clarendon Press, Oxford. pp 229–254Google Scholar
  9. Cock MJ, Sterck L, Rouzé P, Scornet D, Allen AE, Amoutzias G, Anthouard V, Artiguenave F, Aury JM, Badger JH, Beszteri B, Billiau K, Bonnet E, Bothwell JHF, Bowler C, Boyen C, Brownlee C, Carrano CJ, Charrier B, Cho GY, Coelho SM, Collén J, Corre E, Silva CD, Delage L, Delaroque N, Dittami SM, Doulbeau S, Elias M, Farnham G, Gachon CMM, Gschloessl B, Heesch S, Jabbari K, Jubin C, Kawai H, Kimura K, Kloareg B, Küpper FC, Lang D, Le Bail A, Leblanc C, Lerouge P, Lohr M, Lopez PJ, Martens C, Maumus F, Michel G, Miranda-Saavedra D, Morales J, Moreau H, Motomura T, Nagasato C, Napoli CA, Nelson DR, Nyvall-Collén P, Peters AF, Pommier C, Potin P, Poulain J, Quesneville H, Read B, Rensing SA, Ritter A, Rousvoal S, Samanta M, Samson G, Schroeder DC, Ségurens B, Strittmatter M, Tonon T, Tregear J, Valentin L, Von Dassow P, Yamagishi T, Van de Peer Y, Wincker P (2010) The Ectocarpus genome and the independent evolution of multicellularity in the brown algae. Nature 465:617–621PubMedCrossRefGoogle Scholar
  10. Cole DG, Snell WJ (2009) SnapShot: intraflagellar transport. Cell 137(4):784–784PubMedCrossRefGoogle Scholar
  11. Coleman AW (1988) The autofluorescent flagellum: a new phylogenetic enigma. J Phycol 24:118–120CrossRefGoogle Scholar
  12. Fujita S, Iseki M, Yoshikawa S, Makino Y, Watanabe M, Motomura T, Kawai H, Murakami A (2005) Identification and characterization of a fluorescent flagellar protein from the brown alga Scytosiphon lomentaria (Scytosiphonales, Phaeophyceae): a flavoprotein homologous to Old Yellow Enzyme. Eur J Phycol 40:159–167CrossRefGoogle Scholar
  13. Geller A, Müller DG (1981) Analysis of the flagellar beat pattern of male Ectocarpus siliculosus gametes (Phaeophyta) in relation to chemotactic stimulation by female cells. J Exp Biol 92:53–66Google Scholar
  14. Green JC, Leadbeater BSC and Diver WL (1989) The chromophyte algae: problems and perspectives. Oxford Science Publications, Oxford. p. 429Google Scholar
  15. Henry EC, Cole KM (1982a) Ultrastructure of swarmers in the Laminariales (Phaeophyceae). I. Zoospores. J Phycol 18:550–569CrossRefGoogle Scholar
  16. Henry EC, Cole KM (1982b) Ultrastructure of swarmers in the Laminariales (Phaeophyceae). II. Sperms. J Phycol 18:550–569CrossRefGoogle Scholar
  17. Hoyer-Fender S (2010) Centriole maturation and transformation to basal body. Semin Cell Dev Biol 21(2):142–147PubMedCrossRefGoogle Scholar
  18. Kawai H (1988) A flavin-like autofluorescent substance in the posterior flagellum of golden and brown algae. J Phycol 24:114–117CrossRefGoogle Scholar
  19. Kawai H (1992) Green flagellar autofluorescence in brown algal swarmers and their phototactic responses. Bot Mag Tokyo 105:171–184CrossRefGoogle Scholar
  20. Kawai H, Inouye I (1989) Flagellar autofluorescence in forty four chlorophyll c-containing algae. J Phycol 28:222–227CrossRefGoogle Scholar
  21. Kremer J, Mastronarde DN (1996) Computer visualization of three-dimensional image data using IMOD. J Struct Biol 116:71–76PubMedCrossRefGoogle Scholar
  22. La Claire JW II, West JA (1978) Light and electron microscopic studies of growth and reproduction in Cutleria (Phaeophyta). I. Gametogenesis in the female plant of C. hancockii. Protoplasma 97:93–110CrossRefGoogle Scholar
  23. La Claire JW II, West JA (1979) Light and electron microscopic studies of growth and reproduction in Cutleria (Phaeophyta). II. Gametogenesis in the male plant of C. hancockii. Protoplasma 97:93–110CrossRefGoogle Scholar
  24. Lacomble S, Vaughan S, Gadelha C, Morphew MK, Shaw MK, McIntosh JR, Gull K (2009) Three-dimensional cellular architecture of the flagellar pocket and associated cytoskeleton in trypanosomes revealed by electron microscope tomography. J Cell Sci 122:1081–1090PubMedCrossRefGoogle Scholar
  25. Lange BM, Gull K (1995) A molecular marker for centriole maturation in the mammalian cell cycle. J Cell Biol 130:919–927PubMedCrossRefGoogle Scholar
  26. Lofthouse PF, Capon B (1975) Ultrastructural changes accompanying mitosporogenesis in Ectocarpus parvus. Protoplasma 84:83–99PubMedCrossRefGoogle Scholar
  27. Loiseaux S (1973) Ultrastructure of zoidogenesis in unilocular zoidocysts of several brown algae. J Phycol 9:277–289Google Scholar
  28. Loiseaux S, West JA (1970) Brown algal mastigonemes: comparative ultrastructure. Trans Am Microsc Soc 89:524–532CrossRefGoogle Scholar
  29. Maier I (1997a) The fine structure of the male gamete of Ectocarpus siliculosus (Ectocarpales, Phaeophyceae). I. General structure of the cell. Eur J Phycol 32:241–25CrossRefGoogle Scholar
  30. Maier I (1997b) The fine structure of the male gamete of Ectocarpus siliculosus (Ectocarpales, Phaeophyceae). II. The flagellar apparatus. Eur J Phycol 32:255–266CrossRefGoogle Scholar
  31. Manton I (1964) A contribution towards understanding of ‘the primitive fucoid’. New Phytol 63:244–254CrossRefGoogle Scholar
  32. Manton I, Clarke B (1950) Electron microscope observation on the spermatozoids of Fucus. Nature 166:973–974PubMedCrossRefGoogle Scholar
  33. Manton I, Clarke B (1951) Electron microscope observations on the zoospores of Pylaiella and Laminaria. J Exp Bot 2:242–243CrossRefGoogle Scholar
  34. Manton I, Clarke B (1956) Observations with the electron microscope on the internal structure of the spermatozoids of Fucus. J Exp Bot 7:416–329CrossRefGoogle Scholar
  35. Manton I, Clarke B, Greenwood AD (1953) Further observations with the electron microscope on spermatozoids in the brown algae. J Exp Bot 4:319–329CrossRefGoogle Scholar
  36. Markey DR, Bouck GB (1977) Mastigoneme attachment in Ochromonas. J Ultrastruct Res 59:173–177PubMedCrossRefGoogle Scholar
  37. Markey DR, Wilce RT (1975) The ultrastructure of reproduction in the brown alga Pylaiella littoralis. I. Mitosis and cytokinesis in the plurilocular gametangia. Protoplasma 85:219–241PubMedCrossRefGoogle Scholar
  38. Markey DR, Wilce RT (1976a) The ultrastructure of reproduction in the brown alga Pylaiella littoralis. II. Zoosporogenesis in the unilocular sporangia. Protoplasma 88:147–173CrossRefGoogle Scholar
  39. Markey DR, Wilce RT (1976b) The ultrastructure of reproduction in the brown alga Pylaiella littoralis. III. Later stages of gametogenesis in the plurilocular gamtangia. Protoplasma 88:175–186CrossRefGoogle Scholar
  40. Marshall WF (2002) Size control in dynamic organelles. Trends Cell Biol 12(9):414–419PubMedCrossRefGoogle Scholar
  41. Mastronarde DN (1997) Dual-axis tomography: an approach with alignment methods that preserve resolution. J Struct Biol 120:343–352PubMedCrossRefGoogle Scholar
  42. Matsunaga S, Uchida H, IsekiM WM, Murakami A (2010) Flagellar motions in phototactic steering in a brown algal swarmer. Photochem Photobiol 86:374–381PubMedCrossRefGoogle Scholar
  43. McIntosh R, Nicastro D, Mastronarde D (2005) New views of cells in 3D: an introduction to electron tomography. Trends Cell Biol 15:43–51PubMedCrossRefGoogle Scholar
  44. Melkonian M, Reize IB, Preisig HR (1987) Maturation of a flagellum/basal body requires more than one cell cycle in algal flagellates: studies on Nephroselmis olivacea (Prasinophyceae). In: Wiessner W, Robinson DG, Star RC (eds) Algal development. Springer, Berlin, pp 102–103CrossRefGoogle Scholar
  45. Motomura T (1993) Ultrastructural and immunofluorescence studies of zoosporogenesis in Laminaria angustata. Sci Pap Inst Alg Res Fac Sci Hokkaido Univ 9:1–32Google Scholar
  46. Müller D, Falk H (1973) Flagellar structure of the gametes of Ectocarpus siticulosus (Phaeophyta) as revealed by negative staining. Arch Mikrobiol 91:313–322CrossRefGoogle Scholar
  47. Müller DG, Maier I, Müller H (1987) Flagellum autofluorescence and photoaccumulation in heterokont algae. Photochem Photobiol 46(6):1003–1008CrossRefGoogle Scholar
  48. Nagasato C, Motomura T (2002) Influence of the centrosome in cytokinesis of brown algae: polyspermic zygotes of Scytosiphon lomentaria (Scytosiphonales, Phaeophyceae). J Cell Sci 115:2541–2548PubMedGoogle Scholar
  49. Nicastro D (2006) The molecular architecture of axonemes revealed by cryoelectron tomography. Science 313:944–948PubMedCrossRefGoogle Scholar
  50. Nicastro D, McIntosh JR, Baumeister W (2005) 3D structure of eukaryotic flagella in a quiescent state revealed by cryo-electron tomography. Proc Natl Acad Sci USA 102:15889–15894PubMedCrossRefGoogle Scholar
  51. O’Kelly CJ (1989) The evolutionary origin of the brown algae: information from studies of motile cell ultrastructure. In: Green JC, Leadbeater BSC and Diver WL (eds) The chromophyte alge: problems and perspectives. Oxford University Press, Oxford. pp. 255–278Google Scholar
  52. O’Kelly CJ, Floyd GL (1984) The absolute configuration of the flagellar apparatus in zoospores from two species of Laminariales (Phaeophyceae). Protoplasma 123:18–25CrossRefGoogle Scholar
  53. Oda T, Hirokawa N, Kikkawa M (2007) Three-dimensional structures of the flagellar dynein-microtubule complex by cryoelectron microscopy. J Cell Biol 177:243–252PubMedCrossRefGoogle Scholar
  54. Provasoli L (1968) Media and prospects for the cultivation of marine algae. In: Watanabe A, Hattori A (eds) Cultures and collections of algae. Japanese Society of Plant Physiologists, Hakone, pp 63–75Google Scholar
  55. Reynolds ES (1963) The use of lead citrate at high pH as an electron-opaque stain in electronmicroscopy. J Cell Biol 17:208–212PubMedCrossRefGoogle Scholar
  56. Rosenbaum JL, Witman GB (2002) Intraflagellar transport. Nat Rev Mol Cell Biol 3:813–825PubMedCrossRefGoogle Scholar
  57. Schoppmeier J, Lechtreck KF (2003) Flagellar regeneration in Spermatozopsis sililis (Chlorophyta). J Phycol 39:918–922CrossRefGoogle Scholar
  58. Toth R (1974) Sporangial structure and zoosporogenesis in Chorda tomentosa (Laminariales). J Phycol 10:170–185Google Scholar
  59. Toth R, Markey DR (1973) Synaptonemal complexes in brown algae. Nature 243:236–237PubMedCrossRefGoogle Scholar
  60. Ueki C, Nagasto C, Motomura T, Saga N (2008) Reexamination of the pit plugs and the characteristic membranous structures in Porphyra yezoensis (Bangiales, Rhodophyta). Phycol 47:5–11CrossRefGoogle Scholar
  61. Vorobjev IA, Nedezhdina ES (1987) The centrosome and its role in the organization of microtubules. Int Rev Cytol 106:227–229PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Gang Fu
    • 1
    • 2
  • Chikako Nagasato
    • 2
  • Toshiaki Ito
    • 3
  • Dieter G. Müller
    • 4
  • Taizo Motomura
    • 2
    Email author
  1. 1.Graduate School of Environmental ScienceHokkaido UniversitySapporoJapan
  2. 2.Muroran Marine Station, Field Science Center for Northern BiosphereHokkaido UniversityMuroranJapan
  3. 3.Electron Microscope Laboratory, Research Faculty of AgricultureHokkaido UniversitySapporoJapan
  4. 4.Fachbereich BiologieUniversität KonstanzKonstanzGermany

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