A procedure for Dex-induced gene transactivation in Arabidopsis ovules

Abstract

Background Elucidating the genetic and molecular control of plant reproduction often requires the deployment of functional approaches based on reverse or forward genetic screens. The loss-of-function of essential genes, however, may lead to plant lethality prior to reproductive development or to the formation of sterile structures before the organ-of-interest can be analyzed. 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 is otherwise affected by early lethality in conventional, stable mutants.

Arabidopsis CML13 and CML14 Have Essential and Overlapping Roles in Plant Development

“…Among the advantages of the Dex-inducible hpRNAi system is that it facilitates the controlled spatiotemporal suppression of transcripts as transgenic lines can be progressed to homozygosity with low basal expression and good reproducibility (Zhuang et al, 2013;Schubert et al, 2022). This allows for the analysis of potentially essential genes where a complete loss-of-function would otherwise lead to lethality.…”