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Quantitative Evaluation of Plant Actin Cytoskeletal Organization During Immune Signaling

  • Yi-Ju Lu
  • Brad DayEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1578)

Abstract

High spatial and temporal resolution microscopy-based methods are valuable tools for the precise real-time imaging of changes in cellular organization in response to stimulus perception. Here, we describe a quantitative method for the evaluation of the plant actin cytoskeleton during immune stimulus perception and the activation of defense signaling. As a measure of the biotic stress-induced changes in actin filament organization, we present methods for analyzing changes in actin filament organization following elicitation of pattern-triggered immunity and effector-triggered immunity. Using these methods, it is possible to not only quantitatively evaluate changes in actin cytoskeletal organization following biotic stress perception, but to also use these protocols to assess changes in actin filament organization following perception of a wide range of stimuli, including abiotic and developmental cues. As described herein, we present an example application of this method, designed to evaluate changes in actin cytoskeletal organization following pathogen perception and immune signaling.

Key words

Actin cytoskeleton Confocal microscopy Quantitative evaluation ETI PTI Pseudomonas syringae Immune signaling PAMP 

Notes

Acknowledgments

Special thanks to Amy Baetsen-Young for critical review and comments. Work in the Day Lab is supported by a National Science Foundation grant (IOS-1557437).

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Department of Plant, Soil and Microbial SciencesMichigan State UniversityEast LansingUSA
  2. 2.Graduate Program in Cell and Molecular BiologyMichigan State UniversityEast LansingUSA
  3. 3.Graduate Program in GeneticsMichigan State UniversityEast LansingUSA

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