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Transient expression in Arabidopsis thaliana protoplasts derived from rapidly established cell suspension cultures

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Abstract

Arabidopsis thaliana has emerged as a model species for the analysis of genes controlling plant development. However, its small size has impaired biochemical analyses, and the absence of a transient expression system has hampered promoter analysis. Here, we report a method for rapidly establishing A. thaliana suspension cultures that yield protoplasts that can be readily transfected. We have optimized transient expression conditions using a modified polyethylene glycol / calcium nitrate transformation protocol and a Cauliflower Mosaic Virus 35S promoter-luciferase reporter gene construct. Our methods permit isolation of large quantities of rapidly growing cells and analysis of Arabidopsis promoters in vivo in a homologous system.

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Abbreviations

CaMV:

Cauliflower Mosaic Virus

2,4D:

2,4-dichlorophenoxyacetic acid

MES:

2-(N-morpholino)ethanesulfonic acid

PEG:

polyethylene glycol

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Authors and Affiliations

  1. Biology Department and Center for Plant Science and Biotechnology, Washington University, One Brookings Drive, Box 1137, 63130, St. Louis, MO, USA

    Jed H. Doelling & Craig S. Pikaard

Authors
  1. Jed H. Doelling
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  2. Craig S. Pikaard
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Communicated by J. M. Widholm

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Doelling, J.H., Pikaard, C.S. Transient expression in Arabidopsis thaliana protoplasts derived from rapidly established cell suspension cultures. Plant Cell Reports 12, 241–244 (1993). https://doi.org/10.1007/BF00237127

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  • DOI: https://doi.org/10.1007/BF00237127

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