4D Analysis of Arabidopsis Cell Plate Development by Lattice Light Sheet Microscopy

Original video: https://youtu.be/UNe8wa6MpeE

Cell plate formation during cytokinesis entails multiple stages occurring concurrently and requiring orchestrated vesicle delivery, membrane remodelling, and timely deposition of polysaccharides, such as callose. Understanding such a dynamic process requires dissection in time and space; this has been a major hurdle in studying cytokinesis. Using lattice light sheet microscopy (LLSM), we studied cell plate development in four dimensions, through the behavior of yellow fluorescent protein (YFP)-tagged cytokinesis-specific GTPase RABA2a vesicles. We monitored the entire duration of cell plate development, from its first emergence, with the aid of YFP–RABA2a, in both the presence and absence of cytokinetic callose. By developing a robust cytokinetic vesicle volume analysis pipeline, we identified distinct behavioral patterns, allowing the identification of three easily trackable cell plate developmental phases. Notably, the phase transition between phase I and phase II is striking, indicating a switch from membrane accumulation to the recycling of excess membrane material. We interrogated the role of callose using pharmacological inhibition with LLSM and electron microscopy. Loss of callose inhibited the phase transitions, establishing the critical role and timing of the polysaccharide deposition in cell plate expansion and maturation. This study exemplifies the power of combining LLSM with quantitative analysis to decode and untangle such a complex process.

This video compilation provides a comprehensive look at the localization and dynamics of YFP-tagged RABA2a protein at the forming cell plate during plant cell division. High-resolution 4D images were acquired using Light Sheet Microscopy (LLSM), offering a detailed perspective of RABA2a behavior.

Keywords: Callose, cell plate, cytokinesis, 4D imaging, lattice light sheet microscopy, plant cell division, quantitative image analysis, RABA2a

Citation:

Rosalie Sinclair, Minmin Wang, Muhammad Zaki Jawaid, Toshisangba Longkumer, Jesse Aaron, Blair Rossetti, Eric Wait, Kent McDonald, Daniel Cox, John Heddleston, Thomas Wilkop, Georgia Drakakaki, Four-dimensional quantitative analysis of cell plate development in Arabidopsis using lattice light sheet microscopy identifies robust transition points between growth phases, Journal of Experimental Botany, Volume 75, Issue 10, 20 May 2024, Pages 2829–2847, https://doi.org/10.1093/jxb/erae091

Published on: 04 March 2024

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