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.
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
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
Bhandari, N.N. (1984) The microsporangium. In: Embryology of angiosperms, pp. 53–121, Johri, B.M. ed., Springer, Berlin Heidelberg New York Tokyo
Buvat, R. (1989) Ontogeny, cell differentiation, and structure of vascular plants, Springer, Berlin Heidelberg New York
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
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
Dickinson, H.G. (1973) The role of plastids in the formation of pollen grain coatings. Cytobios 8, 25–40
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
Ding, B., Turgeon, R., Parthasarathy, M.V. (1992) Effect of highpressure freezing on plant microfilament bundles. J. Microsc. 165, 367–376
Dobson, H.E.M. (1988) Survey of pollen and pollenkitt lipids — chemical cues to flower visitors? Am. J. Bot. 75, 170–182
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
Edgar, L.A., Pickett-Heaps, J.D. (1982) Ultrastructural localization of polysaccharides in the motile diatom Navicula cuspidata. Protoplasma 113, 10–22
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
Gori, P. (1982) Accumulation of polysaccharides in the anther cavity of Allium sativum, clone Piemonte. J. Ultrastr. Res. 81, 158–162
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
Hastie, R. (1990) Methods in laboratory investigation. Ultrastructure of human basophil leukocytes studied after spray freezing and freeze-substitution. Lab. Invest. 62, 119–130
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
Heslop-Harrison, J. (1968) Tapetal origin of pollen-coat substances in Lilium. New Phytol. 67, 779–786
Hess, M.W. (1990) Spurr resin affects the ultrastructure of freezefixed/freeze-substituted pollen grains. Ultramicroscopy 34, 219–221
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
Hess, M.W., Glaser, A. (1993) A simple and inexpensive device for freeze substitution at 183 K/-90 °C. Biotech. Histochem. 68, 211–214
Hesse, M. (1991) Cytology and morphogenesis of pollen and spores. Prog. Bot. 52, 19–34
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
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
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
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
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
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
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
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
McCauley, M.M., Hepler, P.K. (1992) Cortical ultrastructure of freeze-substituted protonemata of the moss Funaria hygrometrica. Protoplasma 169, 168–178
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
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
Mollenhauer, H.H., Morré, D.J., Griffing, L.R. (1991) Post Golgi apparatus structures and membrane removal in plants. Protoplasma 162, 55–60
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
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
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
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
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
Pacini, E., Franchi, G.G., Hesse, M. (1985) The tapetum: its form, function and possible phylogeny in Embryophyta. Pl. Syst. Evol. 149, 155–185
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
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
Robards, A.W., Stark, M. (1988) Nectar secretion in Abutilon: a new model. Protoplasma 142, 79–91
Rowley, J.R., Dahl, A.O. (1977) Pollen development in Artemisia vulgaris with special reference to glycocalyx material. Pollen Spores 19, 169–184
Schnepf, E. (1969) Über den Feinbau von Öldrüsen. III. Die Ölgänge von Solidago canadensis und die Exkretschläuche von Arctium lappa. Protoplasma 67, 205–212
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
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
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
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
Studer, D., Michel, M., Müller, M. (1989) High-pressure freezing comes of age. Scanning Microsc. Suppl. 3, 253–269
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.
Sütfeld, R., Wiermann, R. (1974) Über die Bedeutung des Antherentapetums für die Akkumulation phenylpropanoider Verbindungen am Pollen. Ber. Deutsch. Bot. Ges. 87, 167–174
Thiéry, J.P. (1967) Mise en évidence des polysaccharides sur coupes fines en microscopie électronique. J. Microscopie 6, 987–1018
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
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.
Wanner, G., Formanek, H., Theimer, R.R. (1981) The ontogeny of lipid bodies (spherosomes) in plant cells. Planta 151, 109–123
Weber, M. (1992) The formation of pollenkitt in Apium nodiflorum (Apiaceae). Ann. Bot. 70, 573–577
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
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
Wiermann, R., Gubatz, S. (1992) Pollen wall and sporopollenin. Int. Rev. Cytol. 140, 35–72
Author information
Authors and Affiliations
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
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
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00198579