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A protocol combining multiphoton microscopy and propidium iodide for deep 3D root meristem imaging in rice: application for the screening and identification of tissue-specific enhancer trap lines

Bureau Charlotte, Lanau Nadège, Ingouff Mathieu, Hassan Boukhaddaoui, Meunier Anne Cecile, Divol Fanchon, Sevilla Rosie, Mieulet Delphine, Diévart Anne, Périn Christophe. 2018. A protocol combining multiphoton microscopy and propidium iodide for deep 3D root meristem imaging in rice: application for the screening and identification of tissue-specific enhancer trap lines. Plant Methods, 14:96, 9 p.

Journal article ; Article de recherche ; Article de revue à facteur d'impact Revue en libre accès total
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Quartile : Q1, Sujet : PLANT SCIENCES / Quartile : Q2, Sujet : BIOCHEMICAL RESEARCH METHODS

Abstract : Background: The clear visualization of 3D organization at the cellular level in plant tissues is needed to fully understand plant development processes. Imaging tools allow the visualization of the main fluorophores and in vivo growth monitoring. Confocal microscopy coupled with the use of propidium iodide (PI) counter-staining is one of the most popular tools used to characterize the structure of root meristems in A. thaliana. However, such an approach is relatively ineffective in species with more complex and thicker root systems. Results: We adapted a PI counter-staining protocol to visualize the internal 3D architecture of rice root meristems using multiphoton microscopy. This protocol is simple and compatible with the main fluorophores (CFP, GFP and mCherry). The efficiency and applicability of this protocol were demonstrated by screening a population of 57 enhancer trap lines. We successfully characterized GFP expression in all of the lines and identified 5 lines with tissue-specific expression. Conclusions: All of these resources are now available for the rice community and represent critical tools for future studies of root development.

Mots-clés Agrovoc : Oryza, Système racinaire, Microscopie immunofluorescente

Classification Agris : F62 - Plant physiology - Growth and development
U30 - Research methods
F50 - Plant structure

Champ stratégique Cirad : Axe 1 (2014-2018) - Agriculture écologiquement intensive

Auteurs et affiliations

  • Bureau Charlotte, CIRAD-BIOS-UMR AGAP (FRA)
  • Lanau Nadège, CIRAD-BIOS-UMR AGAP (FRA)
  • Ingouff Mathieu, UM2 (FRA)
  • Hassan Boukhaddaoui, INSERM (FRA)
  • Meunier Anne Cecile, CIRAD-BIOS-UMR AGAP (FRA)
  • Divol Fanchon, INRA (FRA)
  • Sevilla Rosie, Université de Montpellier (FRA)
  • Mieulet Delphine, CIRAD-BIOS-UMR AGAP (FRA) ORCID: 0000-0001-6220-0372
  • Diévart Anne, CIRAD-BIOS-UMR AGAP (FRA) ORCID: 0000-0001-9460-4638
  • Périn Christophe, CIRAD-BIOS-UMR AGAP (FRA) - auteur correspondant

Source : Cirad-Agritrop (https://agritrop.cirad.fr/589425/)

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