A Procedure for Dex-induced Gene Transactivation in Arabidopsis Ovules

Abstract Background: Elucidating the genetic and molecular control of plant reproduction requires the deployment of functional approaches based, for instance, on reverse or forward genetic screens. The loss-of-function of essential genes, however, may lead to plant lethality prior to reproductive developmentt or to the formation of sterile structures before the organ-of-interest can be analysed. In these cases, inducible approaches that enable a spatial and temporal control of the genetic perturbation are extremely valuable. Genetic induction in reproductive organs, such as the ovule, deeply embedded in the flower, is a delicate procedure that requires both optimization and validation. Results: Here we report on a streamlined procedure enabling reliable induction of gene expression in Arabidopsis ovule and anther tissues using the popular pOP/LhGR Dex-inducible system. We demonstrate its efficiency and reliability using fluorescent reporter proteins and histochemical detection of the GUS reporter gene. Conclusion: The pOP/LhGR system allows for a rapid, efficient and reliable induction of transgenes in developing ovules without compromising developmental progression. This approach opens new possibilities for the functional analysis of candidate regulators in sporogenesis and gametogenesis, which are otherwise affected by early lethality of conventional, stable mutants.