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First published online April 9, 2002; 10.1104/pp.010691

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Plant Physiol, April 2002, Vol. 128, pp. 1390-1401

Pharmacological Analysis of Nod Factor-Induced Calcium Spiking in Medicago truncatula. Evidence for the Requirement of Type IIA Calcium Pumps and Phosphoinositide Signaling1

Eric M. Engstrom, David W. Ehrhardt,2 Raka M. Mitra, and Sharon R. Long*

Howard Hughes Medical Institute, Department of Biological Sciences, Stanford University, Stanford California 94305-5020

Bacterial Nod factors trigger a number of cellular responses in root hairs of compatible legume hosts, which include periodic, transient increases in cytosolic calcium levels, termed calcium spiking. We screened 13 pharmaceutical modulators of eukaryotic signal transduction for effects on Nod factor-induced calcium spiking. The purpose of this screening was 2-fold: to implicate enzymes required for Nod factor-induced calcium spiking in Medicago sp., and to identify inhibitors of calcium spiking suitable for correlating calcium spiking to other Nod factor responses to begin to understand the function of calcium spiking in Nod factor signal transduction. 2-Aminoethoxydiphenylborate, caffeine, cyclopiazonic acid (CPA), 2,5-di-(t-butyl)-1,4-hydroquinone, and U-73122 inhibit Nod factor-induced calcium spiking. CPA and U-73122 are inhibitors of plant type IIA calcium pumps and phospholipase C, respectively, and implicate the requirement for these enzymes in Nod factor-induced calcium spiking. CPA and U-73122 inhibit Nod factor-induced calcium spiking robustly at concentrations with no apparent toxicity to root hairs, making CPA and U-73122 suitable for testing whether calcium spiking is causal to subsequent Nod factor responses.

1 This work was supported by the Howard Hughes Medical Foundation and by the Department of Energy (grant no. DE-FG03-90ER200120). R.M.M. was supported by a Howard Hughes Medical Foundation predoctoral fellowship.

2 Present address: Department of Plant Biology, Carnegie Institution of Washington, Stanford, CA 94305.

* Corresponding author; e-mail srl{at}leland.stanford.edu; fax 650-725-8309.

© 2002 American Society of Plant Physiologists


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