Plant Cell Division Analyzed by Transient Agrobacterium-Mediated Transformation of Tobacco BY-2 Cells

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1370)

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.

Key words

Agrobacterium Transient transformation Tobacco BY-2 Cell division Green fluorescent protein F-actin Cytokinesis 
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Notes

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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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Botany Department, School of Biology and ChemistryOsnabrück UniversityOsnabrückGermany

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