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First published online September 16, 2002; 10.1105/tpc.005579

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The Plant Cell, Vol. 14, 2627-2641, October 2002, Copyright © 2002,
American Society of Plant Biologists

Analysis and Effects of Cytosolic Free Calcium Increases in Response to Elicitors in Nicotiana plumbaginifolia Cells

David Lecourieux1,a, Christian Mazarsb, Nicolas Paulyb, Raoul Ranjevab and Alain Pugin2,a

a Unité Mixte de Recherche Institut National de la Recherche Agronomique–Université de Bourgogne, Biochimie, Biologie Cellulaire et Ecologie des Interactions Plantes-Microorganismes, 17 rue de Sully, BP 86510, 21065 Dijon cedex, France
b Unité Mixte de Recherche Centre National de la Recherche Scientifique–Université Paul Sabatier, Signaux et Messages Cellulaires chez les Végétaux, Pole de Biotechnologies Végétales, 24 chemin de Borde Rouge, BP 17, Auzeville, 31326 Castanet-Tolosan, France

2 To whom correspondence should be addressed. E-mail pugin{at}dijon.inra.fr; fax 03-80-69-32-26

Cell suspensions obtained from Nicotiana plumbaginifolia plants stably expressing the apoaequorin gene were used to analyze changes in cytosolic free calcium concentrations ([Ca2+]cyt) in response to elicitors of plant defenses, particularly cryptogein and oligogalacturonides. The calcium signatures differ in lag time, peak time, intensity, and duration. The intensities of both signatures depend on elicitor concentration and extracellular calcium concentration. Cryptogein signature is characterized by a long-sustained [Ca2+]cyt increase that should be responsible for sustained mitogen-activated protein kinase activation, microtubule depolymerization, defense gene activation, and cell death. The [Ca2+]cyt increase in elicitor-treated cells first results from a calcium influx, which in turns leads to calcium release from internal stores and additional Ca2+ influx. H2O2 resulting from the calcium-dependent activation of the NADPH oxidase also participates in [Ca2+]cyt increase and may activate calcium channels from the plasma membrane. Competition assays with different elicitins demonstrate that [Ca2+]cyt increase is mediated by cryptogein–receptor interaction.




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