Complementation of mutants in plant mitochondrial RNA editing by protoplast transfection

Abstract

A crucial and often decisive test of a nuclear gene being involved in a given process is the complementation of mutants. Restoring the wild type phenotype by the wild type gene introduced into the mutant is a major piece of evidence for the function of this gene. We have developed a rapid and reliable method to complement protoplasts from plants with mutations in mitochondrial RNA editing with the respective wild type genes. The method furthermore allows testing the functionality of modified protein sequences without the need to make and grow transgenic plants, which is very time-consuming. We successfully employed this method for several nuclear-encoded genes involved in RNA editing at specific sites in mitochondria of Arabidopsis thaliana.