Functional transient genetic transformation of Arabidopsis leaves by biolistic bombardment

Shoko Ueki, Benoît Lacroix, Alexander Krichevsky, Sondra G. Lazarowitz, Vitaly Citovsky

Research output: Contribution to journalArticle

62 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)71-77
Number of pages7
JournalNature Protocols
Volume4
Issue number1
DOIs
Publication statusPublished - 2009
Externally publishedYes

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Biolistics
Genetic Transformation
Arabidopsis
Genes
Plant Viral Movement Proteins
Assays
Gene Expression
Agrobacterium
Firearms
Epidermis
Cell Movement
Transcription
Plasmids
Infiltration
Gene expression
Cells
DNA
Tissue
Research

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Functional transient genetic transformation of Arabidopsis leaves by biolistic bombardment. / Ueki, Shoko; Lacroix, Benoît; Krichevsky, Alexander; Lazarowitz, Sondra G.; Citovsky, Vitaly.

In: Nature Protocols, Vol. 4, No. 1, 2009, p. 71-77.

Research output: Contribution to journalArticle

Ueki, Shoko ; Lacroix, Benoît ; Krichevsky, Alexander ; Lazarowitz, Sondra G. ; Citovsky, Vitaly. / Functional transient genetic transformation of Arabidopsis leaves by biolistic bombardment. In: Nature Protocols. 2009 ; Vol. 4, No. 1. pp. 71-77.
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