FIGURE 8.

Characterization of NAE sensitivity and NAE amidohydrolase activity for Arabidopsis seedlings in the presence or absence of PDP-EA or cardanol-EA. A, A. thaliana wild type (Col-0) seedlings with 30 μm NAE 12:0; B, seedlings plus 30 μm NAE 12:0 and 20 μm cardanol-EA; C, seedlings plus 30 μm NAE 12:0 and 20 μm PDP-EA; D, images of the cotyledon sizes of seedlings in presence of 20 μm phenoxyacyl-ethanolamide compounds with or without 30 μm NAE 12:0. Seedlings were grown with these different treatments for 11 days under long day conditions. E and F, sizes of the cotyledons or roots as a function of different NAE, PDP-EA, or cardanol-EA concentrations. G, FAAH activity reactions were initiated by the addition of 6 μg of crude plant protein extract from 11-day-old seedlings in 50 mm BisTris propane-HCl (pH 9.0) at 30 °C for 2 h with 200 μm [1-¹⁴C]NAE 12:0 and 200 μm NAE-like compounds in a final volume of 0.3 ml. Data points represent means ± S.D. (error bars) of triplicate assays. Plots were generated with SigmaPlot software version 12.0. N + C-EA, NAE 12:0 + cardanol-EA. Means with different letters are significantly different (p < 0.005) as determined by one-way analysis of variance with Tukey's post-test.