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.

A procedure for Dex-induced gene transactivation in Arabidopsis ovules / J. Schubert, L. Yanru, M.A. Mendes, D. Fei, H. Dickinson, I. Moore, C. Baroux. - In: PLANT METHODS. - ISSN 1746-4811. - 18:1(2022 Mar 29), pp. 41.1-41.10. [10.1186/s13007-022-00879-x]

A procedure for Dex-induced gene transactivation in Arabidopsis ovules

M.A. Mendes;
2022

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.
English
arabidopsis; dex induction; ovule development; pOP/LhGR
Settore BIO/01 - Botanica Generale
Articolo
Esperti anonimi
Ricerca di base
Pubblicazione scientifica
   PIANO DI SOSTEGNO ALLA RICERCA 2015-2017 - LINEA 2 "DOTAZIONE ANNUALE PER ATTIVITA' ISTITUZIONALE"
   UNIVERSITA' DEGLI STUDI DI MILANO
29-mar-2022
BioMed Central. Part of Springer Nature
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Periodico con rilevanza internazionale
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info:eu-repo/semantics/article
A procedure for Dex-induced gene transactivation in Arabidopsis ovules / J. Schubert, L. Yanru, M.A. Mendes, D. Fei, H. Dickinson, I. Moore, C. Baroux. - In: PLANT METHODS. - ISSN 1746-4811. - 18:1(2022 Mar 29), pp. 41.1-41.10. [10.1186/s13007-022-00879-x]
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J. Schubert, L. Yanru, M.A. Mendes, D. Fei, H. Dickinson, I. Moore, C. Baroux
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/920265
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