Transient Gene Expression in Epidermal Cells of Plant Leaves by Biolistic DNA Delivery

Transient gene expression is a useful approach for studying the functions of gene products. In the case of plants, Agrobacterium in fi ltration is a method of choice for transient introduction of genes for many spe-cies. However, this technique does not work ef fi ciently 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 prob-lems 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 ef fi cient and reproducible transient expression of diverse genes in Arabidopsis, Nicotiana benthami-ana 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