Isolation and Efficient Maize Protoplast Transformation

Abstract

Establishing plant gene function and the study of gene expression regulation often require that genes (or gene variants) are introduced into plants and their activity somehow assayed. Some plants (e.g., Arabidopsis) are easily transformed, while for others (e.g., maize) making stable transgenic plants remains challenging, is expensive and time-consuming. An alternative solution to generating transgenic plants is to assay gene function transiently. In some plants, it is possible to do this by Agroinfiltration. We have adapted a maize protoplast isolation protocol that permits high-efficiency transformation by electroporation, irrespective of the genetic background. Transformed protoplasts have been used to assay gene function using metabolic profiling, to explore protein-DNA interactions using chromatin immunoprecipitation (ChIP), and to assay the activation of reporter constructs by transcription factors. Here, we describe the protocol for efficient maize protoplast isolation and transformation by electroporation.