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Functional transient genetic transformation of Arabidopsis leaves by biolistic bombardment

Nature Protocols volume 4, pages 71–77 (2009)Cite this article

Abstract

Transient gene expression is an indispensable tool for studying functions of gene products. In the case of plants, transient introduction of genes by Agrobacterium infiltration is a method of choice for many species. However, this technique does not work efficiently in Arabidopsis leaf tissue, the most widely used model system for basic plant biology research. Here we present an optimized protocol for biolistic delivery of plasmid DNA into the epidermis of Arabidopsis leaves, which can be easily performed using the Bio-Rad Helios gene gun system. This protocol yields efficient and reproducible transient expression of diverse genes and is exemplified here for use in a functional assay of a transcription repressor and for the subcellular localization and cell-to-cell movement of plant viral movement protein. This protocol is suitable for studies of biological function and subcellular localization of the gene product of interest directly in planta by utilizing different types of activity-based assays. Using this procedure, the data are obtained after 2–4 d of work.

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Figure 1: Arabidopsis plant and leaves suitable for biolistic bombardment.
Figure 2: Transient gene expression to assay transcriptional repressor activity of Arabidopsis SWIRM-domain polyamine oxidase protein 1/lysine demethylase-like protein 1 (SWP1/LDL1) in a transgenic Arabidopsis plant line.
Figure 3: Transient gene expression to assay TMV MP movement in Arabidopsis leaf epidermis.

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Acknowledgements

The work in our laboratory is supported by grants from NIH, NSF, NRI USDA CSREE, BSF and BARD to V.C., and from NIH (1 R01 AI066054-01A) to S.G.L.

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Author notes

  1. Shoko Ueki, Benoît Lacroix and Alexander Krichevsky: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, 11794-5215, New York, USA

    Shoko Ueki, Benoît Lacroix, Alexander Krichevsky & Vitaly Citovsky

  2. Department of Plant Pathology and Plant Microbe Biology, Cornell University, Ithaca, 14853-2703, New York, USA

    Sondra G Lazarowitz

Authors
  1. Shoko Ueki
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  2. Benoît Lacroix
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  3. Alexander Krichevsky
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  4. Sondra G Lazarowitz
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  5. Vitaly Citovsky
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Correspondence to Shoko Ueki.

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Ueki, S., Lacroix, B., Krichevsky, A. et al. Functional transient genetic transformation of Arabidopsis leaves by biolistic bombardment. Nat Protoc 4, 71–77 (2009). https://doi.org/10.1038/nprot.2008.217

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