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A procedure for Dex-induced gene transactivation in Arabidopsis ovules


Schubert, Jasmin; Li, Yanru; Mendes, Marta A; Fei, Danli; Dickinson, Hugh; Moore, Ian; Baroux, Célia (2022). A procedure for Dex-induced gene transactivation in Arabidopsis ovules. Plant Methods, 18:41.

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.

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.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Department of Plant and Microbial Biology
07 Faculty of Science > Zurich-Basel Plant Science Center
Dewey Decimal Classification:580 Plants (Botany)
Scopus Subject Areas:Life Sciences > Biotechnology
Life Sciences > Genetics
Life Sciences > Plant Science
Uncontrolled Keywords:Plant Science, Genetics, Biotechnology
Language:English
Date:2022
Deposited On:13 Apr 2022 09:37
Last Modified:29 Jan 2024 02:39
Publisher:BioMed Central
ISSN:1746-4811
OA Status:Gold
Free access at:PubMed ID. An embargo period may apply.
Publisher DOI:https://doi.org/10.1186/s13007-022-00879-x
PubMed ID:35351175
Project Information:
  • : FunderSNSF
  • : Grant ID310030_185186
  • : Project TitleLinker histones at the nexus of plant cell reprogramming
  • : FunderH2020
  • : Grant ID872417
  • : Project TitleMAD - Mechanisms of Apomictic Developments
  • Content: Published Version
  • Licence: Creative Commons: Attribution 4.0 International (CC BY 4.0)