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Confocal Live Imaging of Shoot Apical Meristems from Different Plant Species

Confocal Live Imaging of Shoot Apical Meristems from Different Plant Species

Published on: March 29, 2019

A protocol combining multiphoton microscopy and propidium iodide for deep 3D root meristem imaging in rice:

Charlotte Bureau¹, Nadège Lanau¹, Mathieu Ingouff²

¹1CIRAD, UMR-AGAP, Université de Montpellier, Avenue Agropolis, 34398 Montpellier Cedex 5, France.

|November 3, 2018

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.

Keywords:

CFP GFP Multiphoton microscope Propidium iodide Rice Root meristem YFP

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