Cell Type-Specific Imaging of Calcium Signaling in Arabidopsis thaliana Seedling Roots Using GCaMP3

Abstract

Cytoplasmic calcium ([Ca²⁺]_cyt) is a well-characterized second messenger in eukaryotic cells. An elevation in [Ca²⁺]_cyt levels is one of the earliest responses in plant cells after exposure to a range of environmental stimuli. Advances in understanding the role of [Ca²⁺]_cyt in plant development has been facilitated by the use of genetically-encoded reporters such as GCaMP. Most of these studies have relied on promoters such as <i>Cauliflower Mosaic Virus</i> (<i>35S</i>) and <i>Ubiquitin10</i> (<i>UBQ10</i>) to drive expression of <i>GCaMP</i> in all cell/tissue types. Plant organs such as roots consist of various cell types that likely exhibit unique [Ca²⁺]_cyt responses to exogenous and endogenous signals. However, few studies have addressed this question. Here, we introduce a set of <i>Arabidopsis thaliana</i> lines expressing <i>GCaMP3</i> in five root cell types including the columella, endodermis, cortex, epidermis, and trichoblasts. We found similarities and differences in the [Ca²⁺]_cyt signature among these root cell types when exposed to adenosine tri-phosphate (ATP), glutamate, aluminum, and salt, which are known to trigger [Ca²⁺]_cyt increases in root cells. These cell type-targeted GCaMP3 lines provide a new resource that should enable more in depth studies that address how a particular environmental stimulus is linked to specific root developmental pathways via [Ca²⁺]_cyt.