Reverse Genetics in Plants

Alain      Tisser; Patrice      Bourgeois

doi:10.2174/1389202013350779

Abstract

Complete genomic sequences of several plant species, most notably the models Arabidopsis thaliana and rice, is now available. One way to discover the biological role of the thousands of new genes is reverse genetics. Plant molecular biologists have already developed several reverse genetics tools. The purpose of this review is to explore the technological avenues taken to address this question and to provide an update on current developments. Because gene targeting by homologous recombination is still not a commodity in plants and despite recent progress with chimeric oligonucleotides, other strategies have been implemented. The most well established routes rely on insertion mutagenesis, either via transposons or Agrobacterium T-DNA transformation. For transposons, except in a few species where highly active endogenous transposable elements exist, this requires the introduction of exogenous elements by transgenesis. The most common strategy for generating insertions relies on binary systems with a st able transposase source and an engineered non-autonomous element. These may contain gene-trap or enhancer-trap devices. This was achieved with both the maize Activator / Dissociation (Ac / Ds) and Enhancer / Inhibitor (En / I) systems in arabidopsis and rice. T-DNA may also be used as an insertion mutagen in species where transformation frequencies are high. In species where transformation is less efficient, gene silencing may prove to be an attractive solution. Finally, the advent of high throughput mutation detection techniques will allow the use of conventional chemically or physically induced mutagenesis in plant reverse genetics. This is theoretically applicable to a wide range of species.

Keywords: genomic sequences, arabidopsis thaliana, actvitor/Dissociation, enhancer inhibitor, arabidopsis, caenorhabditis elegans drosophila, target specificity, Mutation