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Methods Mol Biol. 2017;1578:207-221. doi: 10.1007/978-1-4939-6859-6_17.

Quantitative Evaluation of Plant Actin Cytoskeletal Organization During Immune Signaling.

Lu YJ1, Day B2,3,4.

Author information

1
Department of Plant, Soil and Microbial Sciences, Michigan State University, East Lansing, MI, 48824, USA.
2
Department of Plant, Soil and Microbial Sciences, Michigan State University, East Lansing, MI, 48824, USA. bday@msu.edu.
3
Graduate Program in Cell and Molecular Biology, Michigan State University, East Lansing, MI, 48824, USA. bday@msu.edu.
4
Graduate Program in Genetics, Michigan State University, East Lansing, MI, 48824, USA. bday@msu.edu.

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.

KEYWORDS:

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

PMID:
28220427
DOI:
10.1007/978-1-4939-6859-6_17
[Indexed for MEDLINE]

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