Skip to main content
Log in

Ultrastructural observations on anther tapetum development of freeze-fixed Ledebouria socialis Roth (Hyacinthaceae)

  • Published:
Planta Aims and scope Submit manuscript

Abstract

The tapetal ultrastructure of high-pressure-frozen, freeze-substituted Ledebouria socialis Roth (Hyacinthaceae) is described from the tetrad stage up to microspore mitosis. Cytoplasmic degeneration of the tapeturn occurs after microspore mitosis. During the tetrad stage and the early free-microspore stage the tapetum cells appear to be meristematic; after callose dissolution they show an intense exocytosis of polysaccharides into the anther locule. Later, the tapetum cells are characterized by abundant endoplasmic reticulum (ER). Highly osmiophilic pollenkitt precursor substances accumulate within distinct, partly irregular shaped cytoplasmic domains (“osmiophilic bodies”), which are intimately associated with the ER. It remains to be verified whether or not these bodies are derived from the ER. Because of their preservation and staining patterns the contents of these bodies are tentatively interpreted as flavonoids, one of the main pollenkitt pigments in angiosperms. Apart from these pigment bodies, there exist four other kinds of lipophilic inclusion within the anther (cells). The general aspects of lipid preservation in freeze-substituted samples are discussed. Staining with hot alcoholic phosphotungstic acid yielded good contrast of the ER and other membranes, which are often difficult to visualize in freeze-substituted, resin-embedded samples.

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

E-PTA:

ethanolic phosphotungstic acid

FS:

freeze substitution

HPF:

high-pressure freezing

OBs:

osmiophilic bodies

PA:

periodic acid

PGS:

post-Golgi-apparatus structures

SP:

silver proteinate

TCH:

thiocarbohydrazide

UA:

uranyl acetate

References

  • Albertini, L., Souvré, A., Audran, J.C. (1987) Le tapis de l'anthère et ses relations avec les microsporocytes et les grains de pollen. Rev. Cytol. Biol. végét. Bot. 10, 211–242

    Google Scholar 

  • Beerhues, L., Rittscher, M., Schöpker, H., Schwerdtfeger, C., Wiermann, R. (1993) The significance of the anther tapetum in the biochemistry of pollen pigmentation — an overview. In: The tapetum. Cytology, function, biochemistry and evolution, pp. 117–125 (Pl. Syst. Evol. Suppl. 7), Hesse, M., Pacini, E., Willemse, M.T.M. eds. Springer, Wien New York

    Google Scholar 

  • Bhandari, N.N. (1984) The microsporangium. In: Embryology of angiosperms, pp. 53–121, Johri, B.M. ed., Springer, Berlin Heidelberg New York Tokyo

    Google Scholar 

  • Buvat, R. (1989) Ontogeny, cell differentiation, and structure of vascular plants, Springer, Berlin Heidelberg New York

    Google Scholar 

  • Chandler, D.E. (1984) Comparison of quick-frozen and chemically fixed sea-urchin eggs: structural evidence that cortical granule exocytosis is preceded by a local increase in membrane mobility. J. Cell Sci. 72, 23–36

    Google Scholar 

  • Craig, S., Staehelin, L.A. (1988) High-pressure freezing of intact plant tissues. Evaluation and characterization of novel features of the endoplasmic reticulum and associated membrane systems. Eur. J. Cell Biol. 46, 80–93

    Google Scholar 

  • Dickinson, H.G. (1973) The role of plastids in the formation of pollen grain coatings. Cytobios 8, 25–40

    Google Scholar 

  • Dickinson, H.G., Lewis, D. (1973) The formation of the tryphine coating the pollen grains of Raphanus, and its properties relating to the self-incompatibility system. Proc. R. Soc. Lond. B 184, 149–165

    Google Scholar 

  • Ding, B., Turgeon, R., Parthasarathy, M.V. (1992) Effect of highpressure freezing on plant microfilament bundles. J. Microsc. 165, 367–376

    Google Scholar 

  • Dobson, H.E.M. (1988) Survey of pollen and pollenkitt lipids — chemical cues to flower visitors? Am. J. Bot. 75, 170–182

    Google Scholar 

  • Dumas, C., Charrière-Ladreix, Y. (1979) Etude cytochimique des polyphénols: généralités concernant les lignines et les sécrétions de tannins et d'aglycones flavoniques. Bull. Soc. bot. Fr. 126, Lettres bot. 1979 (2), 123–142

  • Echlin, P. (1971) The role of the tapetum during microsporogenesis of angiosperms. In: Pollen: development and physiology, pp. 41–61, Heslop-Harrison, J., ed. Butterworths, London

    Google Scholar 

  • Edgar, L.A., Pickett-Heaps, J.D. (1982) Ultrastructural localization of polysaccharides in the motile diatom Navicula cuspidata. Protoplasma 113, 10–22

    Google Scholar 

  • Goodwin, T.W. (1965) The biosynthesis of carotenoids. In: Chemistry and biochemistry of plant pigments, pp. 143–174, Goodwin, T.W., ed. Academic Press, London New York

    Google Scholar 

  • Gori, P. (1982) Accumulation of polysaccharides in the anther cavity of Allium sativum, clone Piemonte. J. Ultrastr. Res. 81, 158–162

    Google Scholar 

  • Grant, L, Beversdorf, W.D., Peterson, R.L. (1986) A comparative light and electron microscopic study of microscpore and tapetal development in male fertile and cytoplasmic male sterile oilseed rape (Brassica napus). Can. J. Bot. 64, 1055–1068

    Google Scholar 

  • Hastie, R. (1990) Methods in laboratory investigation. Ultrastructure of human basophil leukocytes studied after spray freezing and freeze-substitution. Lab. Invest. 62, 119–130

    Google Scholar 

  • Herdt, E., Sütfeld, R., Wiermann, R. (1978) The occurrence of enzymes involved in phenylpropanoid metabolism in the tapetum fraction of anthers. Cytobiologie 17, 433–441

    Google Scholar 

  • Heslop-Harrison, J. (1968) Tapetal origin of pollen-coat substances in Lilium. New Phytol. 67, 779–786

    Google Scholar 

  • Hess, M.W. (1990) Spurr resin affects the ultrastructure of freezefixed/freeze-substituted pollen grains. Ultramicroscopy 34, 219–221

    Google Scholar 

  • Hess, M.W. (1992) High-pressure freezing/freeze-substitution of delicate reproductive tissues in plants. In: Electron microscopy 92, vol. III, pp. 67–68, Megías-Megías, L., Rodríguez-García, M.I., Ríos, A., Arias, J.M. eds. Secretariado de Publicaciones de la Universidad de Granada

  • Hess, M.W. (1993) Cell-wall development in freeze-fixed pollen: Inline formation of Ledebouria socialis (Hyacinthaceae). Planta 189, 139–149

    Google Scholar 

  • Hess, M.W., Glaser, A. (1993) A simple and inexpensive device for freeze substitution at 183 K/-90 °C. Biotech. Histochem. 68, 211–214

    Google Scholar 

  • Hesse, M. (1991) Cytology and morphogenesis of pollen and spores. Prog. Bot. 52, 19–34

    Google Scholar 

  • Hesse, M. (1993) Pollenkitt composition in Tilia platyphyllos (Tiliaceae) analyzed by CTEM and EFTEM. In: The tapetum. Cytology, function, biochemistry and evolution, pp. 39–52 (Pl. Syst. Evol. Suppl. 7), Hesse, M., Pacini, E., Willemse, M.T.M., eds. Springer, Wien New York

    Google Scholar 

  • Hoch, H.C. (1991) Preservation of cell ultrastructure by freeze-substitution. In: Electron microscopy of plant pathogens, pp. 1–16, Mendgen, K., Lesemann, D.E., eds. Springer, Berlin Heidelberg New York London Paris Tokyo Hong Kong Barcelona

    Google Scholar 

  • Howard, R.J., Aist, J.R. (1979) Hyphal tip cell ultrastructure of the fungus Fusarium: Improved preservation by freeze-substitution. J. Ultrastr. Res. 66, 224–234

    Google Scholar 

  • Humbel, B., Müller, M. (1986) Freeze substitution and low temperature embedding. In: The science of biological specimen preparation, pp. 175–183, Müller, M., Becker, R.P., Boyde, A., Wolosewick, J.J. eds., SEM Inc., AMF O'Hare, Chicago

    Google Scholar 

  • Locke, M., Krishnan, N. (1971) Hot alcoholic phosphotungstic acid and uranyl acetate as routine stains for thick and thin sections. J. Cell Biol. 50, 550–557

    Google Scholar 

  • Lüttge, U., Schnepf, E. (1976) Organic substances. In: Encyclopedia of Plant Physiology, N. S. vol. 2B: Transports in plants II B. Tissues and organs, pp. 244–277, Lüttge, U., Pitman, M.G., eds. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Mariani, C., de Beuckeleer, M., Truettner, J., Leemans, J., Goldberg, R.B. (1990) Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature 347, 737–741

    Article  CAS  Google Scholar 

  • Marinozzi, V., Luzzatto, A.C., Devirgiliis, L.C., Mazzatenta, C. (1977) Osmium dependent argentaffinity of elastic fibers: its relations with polyphenolic compounds. J. Submicr. Cytol. 9, 267–274

    Google Scholar 

  • McCauley, M.M., Hepler, P.K. (1992) Cortical ultrastructure of freeze-substituted protonemata of the moss Funaria hygrometrica. Protoplasma 169, 168–178

    Google Scholar 

  • Meindl, U., Lancelle, S., Hepler, P.K. (1992) Vesicle production and fusion during lobe formation in Micrasterias visualized by high-pressure freeze fixation. Protoplasma 170, 104–114

    Google Scholar 

  • Mollenhauer, H.H., Totten, C. (1971) Studies on seeds. II. Origin and degradation of lipid vesicles in pea and bean cotyledons. J. Cell Biol. 48, 395–405

    Google Scholar 

  • Mollenhauer, H.H., Morré, D.J., Griffing, L.R. (1991) Post Golgi apparatus structures and membrane removal in plants. Protoplasma 162, 55–60

    Google Scholar 

  • Moor, H. (1987) Theory and practice of high pressure freezing. In: Cryotechniques in biological electron microscopy, pp. 175–191, Steinbrecht, R.A., Zierold, K., eds. Springer, Berlin Heidelberg

    Google Scholar 

  • Murgia, M., Charzynska, M., Rougier, M., Cresti, M. (1991) Secretory tapetum of Brassica oleracea L.: polarity and ultrastructural features. Sex. Plant Reprod. 4, 28–35

    Google Scholar 

  • Nicolas, G. (1991) Advantages of fast-freeze fixation followed by freeze-substitution for the preservation of cell integrity. J. Electr. Microsc. Tech. 18, 395–405

    Google Scholar 

  • Pacini, E. (1990) Tapetum and microspore function. In: Microspores. Evolution and ontogeny, pp. 213–237, Blackmore, S., Knox, R.B., eds. Academic Press, London San Diego New York, Boston Sydney Tokyo Toronto

    Google Scholar 

  • Pacini, E., Franchi, G.G. (1983) Pollen grain development in Smilax aspera L. and possible functions of the loculus. In: Pollen: Biology and implications for plant breeding, pp. 183–190, Mulcahy, D.L., Ottaviano, E., eds. Elsevier Biomedical, New York Amsterdam Oxford

    Google Scholar 

  • Pacini, E., Franchi, G.G., Hesse, M. (1985) The tapetum: its form, function and possible phylogeny in Embryophyta. Pl. Syst. Evol. 149, 155–185

    Google Scholar 

  • Perdue, T.D., Loukides, C.A., Bedinger, P.A. (1992) The formation of cytoplasmic channels between tapetal cells in Zea mays. Protoplasma 171, 75–79

    Google Scholar 

  • Robards, A.W. (1991) Rapid-freezing methods and their application. In: Electron microscopy of plant cells, pp. 257–312, Hall, J.L., Hawes, C., eds. Academic Press, London San Diego New York Boston Sydney Tokyo Toronto

    Google Scholar 

  • Robards, A.W., Stark, M. (1988) Nectar secretion in Abutilon: a new model. Protoplasma 142, 79–91

    Google Scholar 

  • Rowley, J.R., Dahl, A.O. (1977) Pollen development in Artemisia vulgaris with special reference to glycocalyx material. Pollen Spores 19, 169–184

    Google Scholar 

  • Schnepf, E. (1969) Über den Feinbau von Öldrüsen. III. Die Ölgänge von Solidago canadensis und die Exkretschläuche von Arctium lappa. Protoplasma 67, 205–212

    Google Scholar 

  • Seligman, A.M., Wasserkrug, H.L., Hanker, J.S. (1966) A new staining method (OTO) for enhancing contrast of lipid-containing membranes and droplets in osmium tetroxide-fixed tissue with osmiophilic thiocarbohydrazide (TCH). J. Cell Biol. 30, 424–432

    Google Scholar 

  • Staehelin, L.A., Giddings, T.H., Kiss, J.Z., Sack, F.D. (1990) Macromolecular differentiation of Golgi stacks in root tips of Arabidopsis and Nicotiana seedlings as visualized in high-pressure frozen and freeze-substituted samples. Protoplasma 157, 75–91

    Google Scholar 

  • Staiger, D., Kappeler, S., Müller, M., Apel, K. (1993) The proteins encoded by tapetum-specific transcripts Satap35 and Satap44, from Sinapis alba L. are localized in the exine cell wall layer of developing microspores. Planta, in press

  • Stanley, R.G., Linskens, H.F. (1974) Pollen. Biology, biochemistry, managment, Springer, Berlin Heidelberg New York

    Google Scholar 

  • Stein, O., Stein, Y. (1967) Lipid synthesis, intracellular transport, and secretion. II. Electron microscopic radioautograpic study of the mouse lactating mammary gland. J. Cell Biol. 34, 251–263

    Google Scholar 

  • Studer, D., Michel, M., Müller, M. (1989) High-pressure freezing comes of age. Scanning Microsc. Suppl. 3, 253–269

    Google Scholar 

  • Stymne, S., Stobart, A.K. (1987) Triacylglycerol biosynthesis. In: The biochemistry of plants, vol. 9, Lipids: structure and function, pp. 175–214, Stumpf, P.K., ed. Academic Press, Orlando San Diego New York etc.

    Google Scholar 

  • Sütfeld, R., Wiermann, R. (1974) Über die Bedeutung des Antherentapetums für die Akkumulation phenylpropanoider Verbindungen am Pollen. Ber. Deutsch. Bot. Ges. 87, 167–174

    Google Scholar 

  • Thiéry, J.P. (1967) Mise en évidence des polysaccharides sur coupes fines en microscopie électronique. J. Microscopie 6, 987–1018

    Google Scholar 

  • Tiwari, S.C., Gunning, B.E.S. (1986) Development and cell surface of a non-syncytial invasive tapetum in Canna: ultrastructural, freeze-substitution, cytochemical and immunofluorescence study. Protoplasma 134, 1–16

    Google Scholar 

  • van der Meer, M., Stam, M.E., van Tunen, A.J., Mol, J.N.M., Stuitje, A.R. (1992) Inhibition of flavonoid biosynthesis in Petunia anthers by antisense RNA: a novel way to engineer nuclear male sterility. In: Angiosperm pollen and ovules, pp. 22–27, Ottaviano, E., Mulcahy, D.L., Sari-Gorla, M., Bergamini-Mulcahy, G., eds. Springer, New York Berlin Heidelberg etc.

    Google Scholar 

  • Wanner, G., Formanek, H., Theimer, R.R. (1981) The ontogeny of lipid bodies (spherosomes) in plant cells. Planta 151, 109–123

    Google Scholar 

  • Weber, M. (1992) The formation of pollenkitt in Apium nodiflorum (Apiaceae). Ann. Bot. 70, 573–577

    Google Scholar 

  • Weibull, C., Christiansson, A. (1986) Extraction of proteins and membrane lipids during low temperature embedding of biological material for electron microscopy. J. Microsc. 142, 79–86

    Google Scholar 

  • Weibull, C., Villiger, W, Carlemalm, E. (1984) Extraction of lipids during freeze-substitution of Acholeplasma laidlawii-cells for electron microscopy. J. Microsc. 134, 213–216

    Google Scholar 

  • Wiermann, R., Gubatz, S. (1992) Pollen wall and sporopollenin. Int. Rev. Cytol. 140, 35–72

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

We are grateful to Dr. M. Müller (Institut für Zellbiologie, ETH-Zürich, Switzerland) for the kind permission to use the facilities of his laboratory. For helpful discussion, we wish to thank Dr. F. Hadacek, Dr. M. Weber, Mag. M.G. Schlag (Institut für Botanik, Universität Wien, Austria) as well as Dr. M. Müller and Dr. E. Wehrli (Institut für Zellbiologie, ETH-Zürich). We are indebted to Prof. R. Wiermann (Institut für Botanik, Universität Münster, Germany) for critical comments on the manuscript. Thanks are also due to W. Urbancik, A. Glaser and U. Schachner for excellent technical assistence and to the Stadtgärtnerei Zürich for providing the plants. This work was supported by the Austrian Fonds zur Förderung der wissenschaftlichen Forschung (P 8138-BIO). Part of this work was presented at the 8th International Palynological Congress held in September 1992 in Aix-en-Provence/France.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hess, M.W., Hesse, M. Ultrastructural observations on anther tapetum development of freeze-fixed Ledebouria socialis Roth (Hyacinthaceae). Planta 192, 421–430 (1994). https://doi.org/10.1007/BF00198579

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00198579

Key words

Navigation