Electroporation of plant protoplasts and tissues

This chapter discusses the electroporation of plant protoplasts and tissues. Electroporation utilizes short, high-voltage electric shocks to make cells permeable to exogenous molecules. Although direct physical evidence is still lacking, it is widely believed that pores form transiently in the cell membrane in response to these shocks and that the uptake of exogenous molecules occurs through these pores. Electroporation is used to induce the cellular uptake of reporter dyes, enzyme substrates, nucleic acids, and even antibodies and viruses. In plants, electroporation is used primarily to stimulate the uptake of plasmids for stable and transient genetic transformation. Protoplasts are normally utilized in plant electroporation because the cell wall is a major barrier to the diffusion of macromolecules. However, recent work indicates that plasmid DNAs can be introduced into walled plant cells by electroporation. Most of the instruments currently used for electroporation are capacitive-discharge instruments; a capacitor bank is charged by a high-voltage power supply and then discharged into a chamber containing the experimental material. © 1995, Academic Press, Inc.