Transient gene expression in epidermal cells of plant leaves by biolistic DNA delivery

Shoko Ueki, Shimpei Magori, Benoît Lacroix, Vitaly Citovsky

    Research output: Chapter in Book/Report/Conference proceedingChapter

    13 Citations (Scopus)

    Abstract

    Transient gene expression is a useful approach for studying the functions of gene products. In the case of plants, Agrobacterium infiltration is a method of choice for transient introduction of genes for many species. However, this technique does not work efficiently in some species, such as Arabidopsis thaliana. Moreover, the infection of Agrobacterium is known to induce dynamic changes in gene expression patterns in the host plants, possibly affecting the function and localization of the proteins to be tested. These problems can be circumvented by biolistic delivery of the genes of interest. Here, we present an optimized protocol for biolistic delivery of plasmid DNA into epidermal cells of plant leaves, which can be easily performed using the Bio-Rad Helios gene gun system. This protocol allows efficient and reproducible transient expression of diverse genes in Arabidopsis, Nicotiana benthamiana and N. tabacum, and is suitable for studies of the biological function and subcellular localization of the gene products directly in planta. The protocol also can be easily adapted to other species by optimizing the delivery gas pressure.

    Original languageEnglish
    Title of host publicationBiolistic DNA Delivery
    Subtitle of host publicationMethods and Protocols
    EditorsStephan Sudowe, Angelika Reske-Kunz
    Pages17-26
    Number of pages10
    DOIs
    Publication statusPublished - 2013

    Publication series

    NameMethods in Molecular Biology
    Volume940
    ISSN (Print)1064-3745

    Keywords

    • Bio-Rad Helios gene gun system
    • Biolistic gene delivery
    • Leaf epidermis
    • Plant
    • Transient expression

    ASJC Scopus subject areas

    • Molecular Biology
    • Genetics

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