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Published on September 8, 2006; 10.1104/pp.106.085670


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Received June 22, 2006
Accepted August 28, 2006

Extracellular ATP in Plants. Visualization, Localization and Analysis of Physiological Significance in Growth and Signaling

Sung-Yong Kim , Mayandi Sivaguru , and Gary Stacey *

National Center for Soybean Biotechnology and Division of Plant Sciences, University of Missouri-Columbia, Missouri 65211

* Corresponding author; email: staceyg{at}missouri.edu.

Extracellular ATP (eATP) in animals is well documented and known to play an important role in cellular signaling (e.g., at the nerve synapse). The existence of eATP has been postulated in plants; however, there is no definitive experimental evidence for its presence or an explanation as to how such a polar molecule could exit the plant cell and what physiological role it may play in plant growth and development. The presence of eATP in plants (Medicago truncatula) was detected by constructing a novel reporter; fusing a cellulose binding domain peptide to the ATP-requiring enzyme luciferase. Application of this reporter to plant roots allowed the visualization of eATP in the presence of the substrate luciferin. Luciferase activity could be detected in the interstitial spaces between plant epidermal cells and predominantly at the regions of actively growing cells. The levels of eATP were closely correlated with regions of active growth and cell expansion. Pharmacological compounds known to alter cytoplasmic calcium levels, revealed that ATP release is a calcium dependent process and may occur through vesicular fusion; an important step in the polar growth of actively growing root hairs. Reactive oxygen species (ROS) activity at the root hair tip is not only essential for root hair growth but also dependent on the cytoplasmic calcium levels. While application of exogenous ATP and chitin mixture increased ROS activity in root hairs, no changes were observed in response to adenosine, AMP, ADP, and non-hyrolyzable ATP ({beta}{gamma}meATP). However, the application of exogenous Potato apyrase (ATPase) decreased ROS activity suggesting that cytoplasmic calcium gradients and ROS activity are closely associated with eATP release.




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