Abstract
The continuing analysis of plant cell division will require additional protein localization studies. This is greatly aided by GFP-technology, but plant transformation and the maintenance of transgenic lines can present a significant technical bottleneck. In this chapter I describe a method for the Agrobacterium-mediated genetic transformation of tobacco BY-2 cells. The method allows for the microscopic analysis of fluorescence-tagged proteins in dividing cells in within 2 days after starting a coculture. This transient transformation procedure requires only standard laboratory equipment. It is hoped that this rapid method would aid researchers conducting live-cell localization studies in plant mitosis and cytokinesis.
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Acknowledgment
I am grateful to Sabine Zachgo for giving me the possibility to carry out research in the Botany Department, Osnabrück University, Germany. Special thanks to Elison Blancaflor, Noble Foundation, USA, for providing the ABD2-GFP construct in pCAMBIA1390. Many thanks to Clive Lloyd, John Innes Centre, UK, for supporting the development of the transformation technique. This publication was further supported by a grant of the Deutsche Forschungsgemeinschaft (BU 2301/2-1) to HB.
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Buschmann, H. (2016). Plant Cell Division Analyzed by Transient Agrobacterium-Mediated Transformation of Tobacco BY-2 Cells. In: Caillaud, MC. (eds) Plant Cell Division. Methods in Molecular Biology, vol 1370. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3142-2_2
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DOI: https://doi.org/10.1007/978-1-4939-3142-2_2
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3141-5
Online ISBN: 978-1-4939-3142-2
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