Abstract
A key event in the differentiation of elliptically shaped guard cells such as those in Allium is the formation of a radial array of cortical microtubules (Mts) which, by controlling the orientation of wall microfibrils, plays an important role in cell shaping. Previous experiments strongly indicated that the array is nucleated in a zone adjacent to the new ventral wall soon after cytokinesis. In order to further clarify the function of this zone, we performed dual immunolocalizations on Allium guard cells with anti-β-tubulin, to detect Mts, and an antibody to γ-tubulin, a protein known to be present at Mt-organizing centers in other species and recently identified in plants as well. γ-Tubulin antibody stained the cortical zone adjacent to the ventral wall, while little or no fluorescence was present elsewhere along the radial Mt array or at other sites in the cell. The antibody also stained the mitotic poles and phragmoplast in guard mother cells, as it does in other material. No staining was seen when the primary antibody was omitted. The results are consistent with nucleation of the radial array at a cortical-Mt-organizing zone next to the ventral wall, and set the stage for more in-depth studies on the spatial and temporal control of Mt formation in differentiating cells.
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Abbreviations
- CLSM:
-
confocal laser scanning microscope
- FITC:
-
fluorescein isothiocyanate
- Mt:
-
microtubule
- MTOC:
-
microtubule-organizing center
References
Brown, R.C., Lemmon, B.E. (1985) Development of stomata in Selaginella: division polarity and plastid movements. Am. J. Bot. 72, 1914–1925
Busby, C.H., Gunning, B.E.S. (1984) Microtubules and morphogenesis in stomata of the water fern Azolla: an unusual mode of guard cell and pore development. Protoplasma 122, 108–119
Cleary, A.L., Hardham, A.R. (1989) Microtubule organization during development of stomatal complexes in Lolium rigidum. Protoplasma 149, 67–81
Cleary, A.L., Hardham, A.R. (1990) Reinstatement of microtubule arrays from cortical nucleating sites in stomatal complexes of Lolium rigidum following depolymerization of microtubules by oryzalin and high pressure. Plant Cell Physiol. 31, 903–915
Doohan, M.E., Palevitz, B.A. (1980) Microtubules and coated vesicles in guard-cell protoplasts of Allium cepa L. Planta 149, 389–401
Eleftheriou, E.P. (1987) Microtubules and cell wall development in differentiating protophloem sieve elements of Triticum aestivum L. J. Cell Sci. 87, 595–607
Fosket, D.E., Morejohn, L.C. (1992) Structural and functional organization of tubulin. Annu. Rev. Plant Physiol. Plant Mol. Biol. 43, 201–240
Galatis, B. (1980) Microtubules and guard-cell morphogenesis in Zea mays. Protoplasma 45, 211–244
Galatis, B., Apostolakos, P. (1991) Microtubule organization and morphogenesis of stomata in caffeine-treated seedlings of Zea mays. Protoplasma 165, 11–26
Galatis, B., Mitrakos, K. (1980) The ultrastructural cytology of the differentiating guard cells of Vigna sinensis. Am. J. Bot. 67, 1243–1261
Galatis, B., Apostolakos, P., Katsaros, C. (1983) Microtubules and their organizing centres in differentiating guard cells of Adiantum cappilus veneris. Protoplasma 115, 176–192
Hepler, P.K., Palevitz, B.A. (1974) Microtubules and microfilaments. Annu. Rev. Plant Physiol. 25, 309–362
Horio, T., Uzawa, S., Jung, M.K., Oakley, B.R., Tanaka, K., Yanagida, M. (1991) The fission yeast γ-tubulin is essential for mitosis and is located at microtubule organizing centers. J. Cell Sci. 99, 693–700
Joshi, H.C., Palacios, M.J., McNamara, L., Cleveland, D.W. (1992) γ-Tubulin is a centrosomal protein required for cell cycle-dependent microtubule nucleation. Nature 356, 80–83
Jung, G., Wernicke, W. (1990) Cell shaping and microtubules in developing mesophyll of wheat (Triticum aestivum L.). Protoplasma 153, 141–148
Liu, B., Marc, J., Joshi, H.C., Palevitz, B.A. (1993) A γ-tubulin-related protein associated with the microtubule arrays of higher plants in a cell cycle-dependent manner. J. Cell Sci. 104, 1217–1228
Marc, J., Hackett, W.P. (1989) A new method for immunofluorescent localization of microtubules in surface cell layers: application to the shoot apical meristem of Hedera. Protoplasma 148, 70–79
Marc, J., Palevitz, B.A. (1990) Regulation of the spatial order of cortical microtubules in developing guard cells of Allium. Planta 182, 626–634
Marc, J., Mineyuki, Y., Palevitz, B.A. (1989a) The generation and consolidation of a radial array of cortical microtubules in developing guard cells of Allium cepa L. Planta 179, 516–529
Marc, J., Mineyuki, Y., Palevitz, B.A. (1989b) A planar microtubuleorganising center in guard cells of Allium: experimental depolymerisation and reassembly of microtubules. Planta 179, 530–540
Masuda, H., Sevic, M., Cande, W.Z. (1992) In vitro microtubule-nucleating activity of spindle pole bodies in fission yeast Schizosaccharomyces pombe: cell cycle-dependent activation in Xenopus cell-free extracts. J. Cell Biol. 117, 1055–1066
Meagher, R.B. (1991) Divergence and differential expression of actin gene families in higher plants. Int. Rev. Cytol. 125, 139–163
Oakley, B.R. (1992) γ-Tubulin: the microtubule organizer. Trends Cell Biol. 2, 1–5
Oakley, C.E., Oakley, B.R. (1989) The identification of γ-tubulin, a new member of the tubulin superfamily encoded by mip A gene of Aspergillus nidulans. Nature 338, 662–664
Oakley, B.R., Oakley, C.E., Yoon, Y, Jung, M.K. (1990) γ-Tubulin is a component of the spindle pole body that is essential for microtubule function in Aspergillus nidulans. Cell 61, 1289–1301
Palevitz, B.A. (1981) Microtubules and possible microtubule nucleation centers in the cortex of stomatal cells as visualized by high voltage electron microscopy. Protoplasma 107, 115–125
Palevitz, B.A. (1982) The stomatal complex as a model of cytoskeletal participation in cell differentiation. In: The cytoskeleton in plant growth and development, pp. 345–376, Lloyd, C.W., ed. Academic Press, London
Palevitz, B.A., Hepler, P.K. (1976) Cellulose microfibril orientation and cell shaping in developing guard cells of Allium: the role of microtubules and ion accumulation. Planta 132, 71–93
Palevitz, B.A., Mullinax, J.B. (1989) Developmental changes in the arrangement of cortical microtubules in stomatal cells of oat (Avena sativa L.). Cell Motil. Cytoskel. 13, 170–180
Panteris, E., Apostolakos, P., Galatis, B. (1993). Microtubule organization, mesophyll cell morphogenesis, and intercellular space formation in Adiantum cappilus veneris leaflets. Protoplasma 172, 97–110
Pickett-Heaps, J.D. (1969) The evolution of the mitotic apparatus: an attempt at comparative ultrastructural cytology in dividing plant cells. Cytobios 3, 257–280
Quader, H., Deichgraber, G., Schnepf, E. (1986) The cytoskeleton of Cobaea seed hairs: patterning during cell-wall differentiation. Planta 168, 1–10
Sack, F.D., Paolillo, D.J. (1983) Protoplasmic changes during stomatal development in Funaria. Can. J. Bot. 61, 2515–2526
Seagull, R.W., Falconer, M.M. (1991) In vitro xylogenesis. In: The cytoskeletal basis of plant growth and form, pp. 183–194, Lloyd, C.W., ed. Academic Press, London
Stearns, T., Evans, L., Kirschner, M. (1991) γ-Tubulin is a highly conserved element of the centrosome. Cell 65, 825–836
Zheng, Y, Jung, M.K., Oakley, B.R. (1991) γ-Tubulin is present in Drosophila melanogaster and Homo sapiens and is associated with the centrosome. Cell 65, 817–823
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This work was supported by National Science Foundation grant DCB-9019285 to B.A.P., National Institutes of Health (NS30009) and American Cancer Society (CD6255) grants to H.C.J., and a University of Georgia Graduate School Assistantship to B.L. We thank Dr. Mark Farmer and the University of Georgia Center for Advanced Ultrastructural Research for the use of the confocal microscope.
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McDonald, A.R., Liu, B., Joshi, H.C. et al. γ-Tubulin is associated with a cortical-microtubule-organizing zone in the developing guard cells of Allium cepa L.. Planta 191, 357–361 (1993). https://doi.org/10.1007/BF00195693
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DOI: https://doi.org/10.1007/BF00195693