Skip to main content
SpringerLink
Log in

Immunocytochemical localization of photosystem I and the fucoxanthin-chlorophylla/c light-harvesting complex in the diatomPhaeodactylum tricornutum

  • Published:
Protoplasma Aims and scope Submit manuscript

Summary

iserum against two polypeptides of the major fucoxanthin-chlorophylla/c light-harvesting complex of the diatomPhaeodactylum tricornutum and heterologous antiserum against purified photosystem I particles of maize were used to localize these two complexes on the thylakoid membranes ofP. tricornutum. As in many chromophyte algae, the thylakoids are loosely appressed and organized into extended bands of three, giving a ratio of 2∶1 for appressed versus non-appressed membranes. Immunoelectron microscopy demonstrated that the fucoxanthin-chlorophylla/c light-harvesting complex, which is believed to be associated with photosystem II, was equally distributed on the appressed and non-appressed thylakoid membranes. Photosystem I was also found on both types of membranes, but was slightly more concentrated on the two outer non-appressed membranes of each band. Similarly, photosystem I activity, as measured by the photooxidation of 3,3′-diaminobenzidine, was higher in the outer thylakoids than in the central thylakoid of each band. We conclude that the thylakoids of diatoms differ from those of green algae and higher plants in their macromolecular organization as well as in their morphological arrangement.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BSA:

bovine serum albumin

DAB:

3,3′-diaminobenzidine

FCPC:

fucoxanthin-chlorophylla/c light-harvesting complex

LHC:

light-harvesting complex

PBS:

phosphate-buffered saline

PS:

photosystem

References

  • Allred DR, Staehelin LA (1985) Lateral distribution of the cytochrome b6/f and coupling factor ATP synthetase complexes of chloroplast thylakoid membranes. Plant Physiol 78: 199–202

    Google Scholar 

  • — — (1986) Spatial organization of the cytochrome b6/f complex within chloroplast thylakoid membranes. Biochim Biophys Acta 849: 94–103

    Google Scholar 

  • Anderson JM, Andersson B (1982) The architecture of photosynthetic membranes: lateral and transverse organization. Trends Biochem Sci 7: 288–292

    Google Scholar 

  • Andersson B, Anderson JM (1980) Lateral heterogeneity in the distribution of chlorophyll-protein complexes of the thylakoid membranes of spinach chloroplasts. Biochim Biophys Acta 593: 427–440

    Google Scholar 

  • Bennett J (1983) Regulation of photosynthesis by reversible phosphorylation of the light-harvesting chlorophylla/b protein. Biochem J 212: 1–13

    Google Scholar 

  • Berkaloff C, Duval JC, Hauswirth N, Rousseau B (1983) Freeze fracture study of thylakoids ofFucus serratus. J Phycol 19: 96–100

    Google Scholar 

  • Callahan FE, Wergin WP, Nelson N, Edelman M, Mattoo AK (1989) Distribution of thylakoid proteins between stromal and granal lamellae inSpirodela: dual location of photosystem II components. Plant Physiol 91: 629–635

    Google Scholar 

  • Cox RP, Andersson B (1981) Lateral and transverse organisation of cytochromes in the chloroplast thylakoid membrane. Biochem Biophys Res Commun 103: 1336–1342

    Google Scholar 

  • Dwarte DM, Vesk M (1982) Freeze-fracture thylakoid ultrastructure of representative members of “chlorophyll c” algae. Micron 13: 325–326

    Google Scholar 

  • — — (1983) A freeze-fracture study of cryptomonad thylakoids. Protoplasma 117: 130–141

    Google Scholar 

  • Fawley MW, Grossman AR (1986) Polypeptides of a light-harvesting complex of the diatomPhaeodactylum tricornutum are synthesized in the cytoplasm of the cell as precursors. Plant Physiol 81: 149–155

    Google Scholar 

  • Frederick SE (1987) DAB procedures. In: Vaughn KC (ed) CRC handbook of plant cytochemistry, vol 1. CRC Press, Boca Raton, FL, pp 3–23

    Google Scholar 

  • Goodchild DJ, Anderson JM, Andersson B (1985) Immunocytochemical localization of the cytochrome b/f complex of chloroplast thylakoid membranes. Cell Biol Int Rep 9: 715–721

    Google Scholar 

  • Herman EM (1988) Immunocytochemical localization of macro-molecules with the electron microscope. Annu Rev Plant Phys Plant Mol Biol 39: 139–155

    Google Scholar 

  • Lichtlé C, Duval JC, Hauswirth N, Spilar A (1986) Freeze fracture study of thylakoid organization ofCryptomonas rufescens (Cryptophyceae) according to illumination conditions. Photobiochem Photobiophys 11: 159–171

    Google Scholar 

  • - McKay RML, Gibbs SP (1991) Immunogold localization of PS I and PS II light-harvesting complexes in cryptomonad thylakoids. Biol Cell (submitted)

  • Ludwig M, Gibbs SP (1989) Localization of phycoerythrin at the lumenal surface of the thylakoid membrane inRhodomonas lens. J Cell Biol 108: 875–884

    Google Scholar 

  • Manodori A, Grossman AR (1990) Sequence homology between light harvesting polypeptides of plants and the diatomPhaeodactylum tricornutum. In: Baltscheffsky M (ed) Current research in photosynthesis, vol 3. Kluwer, Hingham, MA, pp 541–544

    Google Scholar 

  • McKay RML, Gibbs SP (1990) Phycoerythrin is absent from the pyrenoid ofPorphyridium cruentum: photosynthetic implications. Planta 180: 249–256

    Google Scholar 

  • — — (1991) Composition and function of pyrenoids: cytochemical and immunocytochemical approaches. Can J Bot 69: 1040–1052

    Google Scholar 

  • McLachlan J (1973) Growth media-marine. In: Stein JR (ed) Handbook of phycological methods. Culture methods and growth measurements. Cambridge University Press, New York, pp 25–51

    Google Scholar 

  • Miller KR, Staehelin LA (1976) Analysis of the thylakoid outer surface: coupling factor is limited to unstacked membrane regions. J Cell Biol 68: 30–47

    Google Scholar 

  • Mullet JE (1983) The amino acid sequence of the polypeptide segment which regulates membrane adhesion (grana stacking) in chloroplasts. J Biol Chem 258: 9941–9948

    Google Scholar 

  • Owens TG (1986) Light-harvesting function in the diatomPhaeodactylum tricornutum. II. Distribution of excitation energy between the photosystems. Plant Physiol 80: 739–746

    Google Scholar 

  • Rhiel E, Mörschel E, Wehrmeyer W (1985) Correlation of pigment deprivation and ultrastructural organization of thylakoid membranes inCryptomonas maculata following nutrient deficiency. Protoplasma 129: 62–73

    Google Scholar 

  • Roth J (1982) The protein A-gold (pAg) technique-a qualitative and quantitative approach for antigen localization on thin sections. In: Bullock GR, Petrusz P (eds) Techniques in immunocytochemistry, vol 1. Academic Press, New York, pp 127–133

    Google Scholar 

  • Shaw PJ, Henwood JA (1985) Immuno-gold localization of cytochrome f, light-harvesting complex, ATP synthase and ribulose 1,5-bisphosphate carboxylase/oxygenase. Planta 165: 333–339

    Google Scholar 

  • Simpson DJ, n Wettstein D (1989) The structure and function of the thylakoid membrane. Carlsberg Res Commun 54: 55–65

    Google Scholar 

  • Slot JW, Geuze HJ (1981) Sizing of protein A-colloidal gold probes for immunoelectron microscopy. J Cell Biol 90: 533–536

    Google Scholar 

  • Smith BM, Melis A (1988) Photochemical apparatus organization in the diatomCylindrotheca fusiformis: photosystem stoichiometry and excitation distribution in cells grown under high and low irradiance. Plant Cell Physiol 29: 761–769

    Google Scholar 

  • Spear-Bernstein L, Miller KR (1985) Are the photosynthetic membranes of cryptophyte algae inside out? Protoplasma 129: 1–9

    Google Scholar 

  • Spurr AR (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26: 31–43

    Google Scholar 

  • Staehelin LA (1986) Chloroplast structure and supramolecular organization of photosynthetic membranes. In: Staehelin LA, Arntzen CJ (eds) Photosynthesis III. Springer, Berlin Heidelberg New York Tokyo, pp 1–84 [Pirson A, Zimmerman MH (eds) Encyclopedia of plant physiology, NS, vol 19]

    Google Scholar 

  • —, Arntzen CJ (1983) Regulation of chloroplast membrane function: protein phosphorylation changes the spatial organization of membrane components. J Cell Biol 97: 1327–1337

    Google Scholar 

  • Starr RC, Zeikus JA (1987) UTEX-the culture collection of algae at the University of Texas at Austin. J Phycol 23 s: 1–47

    Google Scholar 

  • Sweeney BM (1981) Freeze-fractured chloroplast membranes ofGonyaulax polyedra (Pyrrophyta). J Phycol 17: 95–101

    Google Scholar 

  • Tschopp J (1984) Ultrastructure of the membrane attack complex of complement. Heterogeneity of the complex caused by different degree of C9 polymerization. J Biol Chem 259: 7857–7863

    Google Scholar 

  • Vallon O, Wollman FA, Olive J (1985) Distribution of intrinsic and extrinsic subunits of the PS II protein complex between appressed and non-appressed regions of the thylakoid membrane: an immunocytochemical study. FEBS Lett 183: 245–250

    Google Scholar 

  • — — — (1986) Lateral distribution of the main protein complexes of the photosynthetic apparatus inChlamydomonas reinhardtii and in spinach: an immunocytochemical study using intact thylakoid membranes and a PS II enriched membrane preparation. Photobiochem Photobiophys 12: 203–220

    Google Scholar 

  • Webber AN, Platt-Aloia KA, Heath RL, Thomson WW (1988) The marginal regions of thylakoid membranes: a partial characterization by polyoxyethylene sorbitan monolaurate (Tween 20) solubilization of spinach thylakoids. Physiol Plant 72: 288–297

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biology, McGill University, 1205 Docteur Penfield Avenue, H3A 1B1, Montreal, Quebec, Canada

    A. M. Pyszniak & Sarah P. Gibbs

Authors
  1. A. M. Pyszniak
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sarah P. Gibbs
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pyszniak, A.M., Gibbs, S.P. Immunocytochemical localization of photosystem I and the fucoxanthin-chlorophylla/c light-harvesting complex in the diatomPhaeodactylum tricornutum . Protoplasma 166, 208–217 (1992). https://doi.org/10.1007/BF01322783

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01322783

Keywords

Search

Navigation