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J Vis Exp

Title:		"Wide-Field, Real-Time Imaging of Local and Systemic Wound Signals in Arabidopsis"
Author(s):		Uemura T; Wang J; Aratani Y; Gilroy S; Toyota M;
Address:		"Department of Biochemistry and Molecular Biology, Saitama University. Department of Botany, University of Wisconsin. Department of Biochemistry and Molecular Biology, Saitama University; Department of Botany, University of Wisconsin; mtoyota@mail.saitama-u.ac.jp"
Journal Title:		J Vis Exp
Year:		2021
Volume:		20210604
Issue:		172
Page Number:		-
DOI:		10.3791/62114
ISSN/ISBN:		1940-087X (Electronic) 1940-087X (Linking)
Abstract:		"Plants respond to mechanical stresses such as wounding and herbivory by inducing defense responses both in the damaged and in the distal undamaged parts. Upon wounding of a leaf, an increase in cytosolic calcium ion concentration (Ca(2+) signal) occurs at the wound site. This signal is rapidly transmitted to undamaged leaves, where defense responses are activated. Our recent research revealed that glutamate leaking from the wounded cells of the leaf into the apoplast around them serves as a wound signal. This glutamate activates glutamate receptor-like Ca(2+) permeable channels, which then leads to long-distance Ca(2+) signal propagation throughout the plant. The spatial and temporal characteristics of these events can be captured with real-time imaging of living plants expressing genetically encoded fluorescent biosensors. Here we introduce a plant-wide, real-time imaging method to monitor the dynamics of both the Ca(2+) signals and changes in apoplastic glutamate that occur in response to wounding. This approach uses a wide-field fluorescence microscope and transgenic Arabidopsis plants expressing Green Fluorescent Protein (GFP)-based Ca(2+) and glutamate biosensors. In addition, we present methodology to easily elicit wound-induced, glutamate-triggered rapid and long-distance Ca(2+) signal propagation. This protocol can also be applied to studies on other plant stresses to help investigate how plant systemic signaling might be involved in their signaling and response networks"
Keywords:		"*Arabidopsis/genetics/metabolism *Arabidopsis Proteins/metabolism Calcium/metabolism Cytosol/metabolism Gene Expression Regulation, Plant Herbivory Plant Leaves/metabolism;"
Notes:		"MedlineUemura, Takuya Wang, Jiaqi Aratani, Yuri Gilroy, Simon Toyota, Masatsugu eng Video-Audio Media 2021/06/22 J Vis Exp. 2021 Jun 4; (172). doi: 10.3791/62114"