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A procedure for Dex-induced gene transactivation in Arabidopsis ovules

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 th...

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Autores principales: Schubert, Jasmin, Li, Yanru, Mendes, Marta A., Fei, Danli, Dickinson, Hugh, Moore, Ian, Baroux, Célia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8962214/
https://www.ncbi.nlm.nih.gov/pubmed/35351175
http://dx.doi.org/10.1186/s13007-022-00879-x
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author Schubert, Jasmin
Li, Yanru
Mendes, Marta A.
Fei, Danli
Dickinson, Hugh
Moore, Ian
Baroux, Célia
author_facet Schubert, Jasmin
Li, Yanru
Mendes, Marta A.
Fei, Danli
Dickinson, Hugh
Moore, Ian
Baroux, Célia
author_sort Schubert, Jasmin
collection PubMed
description 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13007-022-00879-x.
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spelling pubmed-89622142022-03-30 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 Plant Methods Methodology 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13007-022-00879-x. BioMed Central 2022-03-29 /pmc/articles/PMC8962214/ /pubmed/35351175 http://dx.doi.org/10.1186/s13007-022-00879-x Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Methodology
Schubert, Jasmin
Li, Yanru
Mendes, Marta A.
Fei, Danli
Dickinson, Hugh
Moore, Ian
Baroux, Célia
A procedure for Dex-induced gene transactivation in Arabidopsis ovules
title A procedure for Dex-induced gene transactivation in Arabidopsis ovules
title_full A procedure for Dex-induced gene transactivation in Arabidopsis ovules
title_fullStr A procedure for Dex-induced gene transactivation in Arabidopsis ovules
title_full_unstemmed A procedure for Dex-induced gene transactivation in Arabidopsis ovules
title_short A procedure for Dex-induced gene transactivation in Arabidopsis ovules
title_sort procedure for dex-induced gene transactivation in arabidopsis ovules
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8962214/
https://www.ncbi.nlm.nih.gov/pubmed/35351175
http://dx.doi.org/10.1186/s13007-022-00879-x
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