Plant Physiol, April 2003, Vol. 131, pp. 1781-1791

N-Acylethanolamine Signaling in Tobacco Is Mediated by a Membrane-Associated, High-Affinity Binding Protein¹

Department of Biological Sciences, University of North Texas, Denton, Texas 76203 (S.T., K.K.-S., K.D.C.); and Plant Biology Division, The Samuel Roberts Noble Foundation, Ardmore, Oklahoma 73402 (R.A.D.)

N-Acylethanolamines (NAEs) are fatty acid derivatives found as minor constituents of animal and plant tissues, and their levels increase 10- to 50-fold in tobacco (Nicotiana tabacum) leaves treated with fungal elicitors. Infiltration of tobacco leaves with submicromolar to micromolar concentrations of N-myristoylethanolamine (NAE 14:0) resulted in an increase in relative phenylalanine ammonia-lyase (PAL) transcript abundance within 8 h after infiltration, and this PAL activation was reduced after co-infiltration with cannabinoid receptor antagonists (AM 281 and SR 144528). A saturable, high-affinity specific binding activity for [³H]NAE 14:0 was identified in suspension-cultured tobacco cells and in microsomes from tobacco leaves (apparent K_d of 74 and 35 nM, respectively); cannabinoid receptor antagonists reduced or eliminated specific [³H]NAE 14:0 binding, consistent with the physiological response. N-Oleoylethanolamine activated PAL2 expression in leaves and diminished [³H]NAE 14:0 binding in microsomes, whereas N-linoleoylethanolamine did not activate PAL2 expression in leaves, and did not affect [³H]NAE 14:0 binding in microsomes. The nonionic detergent dodecylmaltoside solubilized functional [³H]NAE 14:0-binding activity from tobacco microsomal membranes. The dodecylmaltoside-solubilized NAE-binding activity retained similar, but not identical, binding properties to the NAE-binding protein(s) in intact tobacco microsomes. Additionally, high-affinity saturable NAE-binding proteins were identified in microsomes isolated from Arabidopsis and Medicago truncatula tissues, indicating the general prevalence of these binding proteins in plant membranes. We propose that plants possess an NAE-signaling pathway with functional similarities to the "endocannabinoid" pathway of animal systems and that this pathway, in part, participates in xylanase elicitor perception in tobacco.