Summary
Immunofluorescence microscopy using an antibody against xyloglucan (XG) revealed its dynamics during the cell cycle. In interphase tobacco BY-2 cells, punctate and scattered fluorescence was observed throughout the cytoplasm. Colocalization of such signals with cortical microtubules (MTs) was clearly observed on the membrane ghosts. They were also associated and accumulated on MT bundles of the preprophase band. Treatment of protoplasts with cytochalasin B prior to the preparation of the ghosts had no effect on the pattern of anti-XG staining, while treatment with propyzamide caused the disappearance of the staining. These results suggest an association of Golgi apparatus and/or Golgi-derived vesicles with MTs. In metaphase cells, the staining was dispersed in the cytoplasm, except in the area occupied by the metaphase spindle. During anaphase, a broad fluorescence band appeared between daughter chromosomes and gradually concentrated at the equatorial plane before formation of the phragmoplast. At telophase, a bright line of fluorescence appeared at the equatorial plane corresponding to the position of the cell plate. The length of the line increased as cytokinesis proceeded. Thus, we showed that immunofluorescence microscopy using anti-XG antibody can be considered as a powerful tool for the analysis of Golgi apparatus and Golgi-derived vesicles containing XG.
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References
Baba K, Sone Y, Misaki A, Hayashi T (1994) Localization of xyloglucan in the macromolecular complex composed of xyloglucan and cellulose in pea stems. Plant Cell Physiol 35: 439–444
Bajer A (1965) Cine micrographie analysis of cell plate formation in endosperm. Exp Cell Res 37: 376–398
Baskin TI, Cande WZ (1990) The structure and function of the mitotic spindle in flowering plants. Annu Rev Plant Physiol 41: 277–315
Dute RR, Rushing AE (1988) Notes on torus development in the wood ofOsmanthus americanus (L) Benth. & Hook. ex Gray (Oleaceae). IAWA Bull NS 9: 41–51
— —, Perry JW (1990) Torus structure and development in species ofDaphne, IAWA Bull NS 11: 401–412
Gu X, Verma DP (1996) Phragmoplastin, a dynamin-like protein associated with cell plate formation in plants. EMBO J 15: 695–704
Gunning BES (1982) The cytokinetic apparatus: its development and spatial regulation. In: Lloyd CW (ed) The cytoskeleton in plant growth and development. Academic Press, London, pp 229–292
Hayashi T (1989) Xyloglucans in the primary cell wall. Annu Rev Plant Physiol Plant Mol Biol 40: 139–168
Hogetsu T (1991) Mechanism for formation of the secondary wall thickening in tracheary elements: microtubules and microfibrils of tracheary elements ofPisum sativum L. andCommelina communis L. and the effects of amiprophosmethyl. Planta 185: 190–200
Kakimoto T, Shibaoka H (1988) Cytoskeletal ultrastructure of phragmoplast-nuclei complexes isolated from cultured tobacco cells. Protoplasma Suppl 2: 95–103
— — (1992) Synthesis of polysaccharides in phragmoplasts isolated from tobacco BY-2 cells. Plant Cell Physiol 33: 353–361
Kawazu T, Kawano S, Kuroiwa T (1995) Distribution of the Golgi apparatus in the mitosis of cultured tobacco cells as revealed by DiOC6 fluorescence microscopy. Protoplasma 186: 183–192
Lauber MH, Waizenegger I, Steinmann T, Schwarz H, Mayer U, Hwang I, Lukowitz W, Jurgens G (1997) The Arabidopsis KNOLLE protein is a cytokinesis-specific syntaxin. J Cell Biol 139: 1485–1493
Moore PJ, Staehelin LA (1988) Immunogold localization of the cell-wall-matrix polysaccharides rhamnogalacturonan I and xyloglucan during cell expansion and cytokinesis inTrifolium pratense L.: implication for secretory pathways. Planta 174: 433–445
—, Darvill AG, Albersheim P, Staehelin LA (1986) Immunogold localization of xyloglucan and rhamnogalacturonan I in the cell walls of suspension-cultured Sycamore cells. Plant Physiol 82: 787–794
Nagata T, Okada K, Takebe I, Mitsui C (1981) Delivery of tobacco mosaic virus RNA into plant protoplasts mediated by reverse-phase evaporation vesicles (liposomes). Mol Gen Genet 184: 161–165
—, Okada K, Takebe I (1982) Mitotic protoplasts and their infection with tobacco mosaic virus RNA encapsulated in liposomes. Plant Cell Rep 1: 250–252
—, Nemoto Y, Hasezawa S (1992) Tobacco BY-2 cell line as the “HeLa” cells in the cell biology of higher plants. Int Rev Cytol 132: 1–30
Northcote DH, Davey R, Lay J (1989) Use of antisera to localize callose, xylan and arabinogalactan in the cell-plate, primary and secondary walls of plant cells. Planta 178: 353–366
Park JM, Kang SG, Pih KT, Jang HJ, Piao HL, Yoon HW, Cho MJ, Hwang I (1997) A dynamin-like protein, ADL1, is present in membranes as a high-molecular-mass complex inArabidopsis thaliana. Plant Physiol 115: 763–771
Pickett-Heaps JD (1967) Further observations on the Golgi apparatus and its functions in cells of the wheat seedling. J Ultrastruct Res 18: 287–303
Samuels AL, Giddings TH Jr, Staehelin LA (1995) Cytokinesis in tobacco BY-2 and root tip cells: a new model of cell plate formation in higher plants. J Cell Biol 130: 1345–1357
Shibaoka H, Asada T, Yamamoto S, Sonobe S (1996) The use of model systems prepared from tobacco BY-2 cells for studies of the plant cytoskeleton. J Microsc 181: 145–152
Sone Y, Fujikawa Y (1993) Estimation of xyloglucan in vegetables by enzyme-linked immunosorbent assay. J Nutr Sci Vitaminol 39: 597–606
—, Kuramae J, Shibata S, Missaki A (1989) Immunochemical specificities of antibody to the heptasaccharide unit of plant xyloglucan. Agric Biol Chem 53: 2821–2823
Sonobe S, Takahashi S (1994) Association of microtubules with the plasma membrane of tobacco BY-2 cells in vitro. Plant Cell Physiol 35: 451–460
Staehelin LA, Hepler PK (1996) Cytokinesis in higher plants. Cell 84: 821–824
Whaley WG, Mollenhauer HH (1963) The Golgi apparatus and cell plate formation: a postulate. J Cell Biol 17: 216–221
Wick SM (1991) The preprophase band. In: Lloyd CW (ed) The cytoskeletal basis of plant growth and form. Academic Press, London, pp 231–244
Yasuhara H, Sonobe S, Shibaoka H (1993) Effects of taxol on the development of the cell plate and of the phragmoplast in tobacco BY-2 cells. Plant Cell Physiol 34: 21–29
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Sonobe, S., Nakayama, N., Shimmen, T. et al. Intracellular distribution of subcellular organelles revealed by antibody against xyloglucan during cell cycle in tobacco BY-2 cells. Protoplasma 213, 218–227 (2000). https://doi.org/10.1007/BF01282159
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DOI: https://doi.org/10.1007/BF01282159