Summary
Tobacco BY-2 cells have become a major tool in plant cell biological research, in part due to the availability of a cell cycle synchronization protocol. This method, pioneered by Nagata and coworkers, involves sequential treatments with aphidicolin (a DNA synthesis inhibitor) and propyzamide (a microtubule inhibitor which arrests mitosis). The effects of these inhibitors are reversible, allowing the cell culture to progress into M phase synchronously. However, attempts to reproduce high levels of synchrony with published protocols have not been uniformly successful. This paper describes critical parameters for cell cycle synchronization and documents the kinetics and variation typically found in using this protocol.
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References
Akashi T, Izumi K, Nagano M, Enomoto E, Mizuno K, Shibaoka H (1988) Effects of propyzamide on tobacco cell microtubules in vivo and in vitro. Plant Cell Physiol 29: 1053–1062
Asada T, Shibaoka H (1994) Isolation of polypeptides with microtubule-translocating activity from phragmoplasts of tobacco BY-2 cells. J Cell Sci 107: 2249–2257
—, Sonobe S, Shiboaka H (1991) Microtubule translocation in the cytokinetic apparatus of cultured tobacco cells. Nature 350: 238–241
Hasezawa S, Sano T, Nagata T (1994) Oblique cell plate formation in tobacco BY-2 cells originates in double preprophase bands. J Plant Res 107: 355–359
Kakimoto T, Shibaoka H (1988) Cytoskeletal ultrastructure of phragmoplast-nuclei complexes isolated from cultured tobacco cells. Protoplasma Suppl 2: 95–103
Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18: 100–127
Nagata T, 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” cell in the cell biology of higher plants. Int Rev Cytol 132: 1–30
Sala F, Parisi B, Burroni D, Amileni AR, Pedrali-Noy G, Spadari S (1980) Specific and reversible inhibition by aphidicolin of the a-like DNA polymerase of plant cells. FEBS Lett 117: 93–98
Samuels AL, Staehelin LA (1996) Caffeine inhibits cell plate formation by disrupting membrane reorganization just after the vesicle fusion step. Protoplasma 195: 144–155
—, 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
Spadari S, Sala F, Pedrali-Noy G (1982) Aphidicolin: a specific inhibitor of nuclear DNA replication in eukaryotes. Trends Biochem Sci 62: 29–32
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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Samuels, A.L., Meehl, J., Lipe, M. et al. Optimizing conditions for tobacco BY-2 cell cycle synchronization. Protoplasma 202, 232–236 (1998). https://doi.org/10.1007/BF01282551
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DOI: https://doi.org/10.1007/BF01282551