Levels of phytohormones increase in Populus nigra leaves over the course of rust fungus infection. Hormone metabolites were analyzed in leaves of rust infected (closed circles) and noninfected control (open circles) trees at different time points after inoculation. Samples were collected from separate trees at each time point. Concentration of (a) salicylic acid (SA), (b) salicylic acid glucoside (SAG), (c) abscisic acid (ABA), (d) jasmonic acid (JA), (e) jasmonic acid glucoside (JA‐Glc), and (f) jasmonic acid‐isoleucine (JA‐Ile). Data were analyzed by two‐way ANOVA. Data are presented as the mean ± SE (n = 5), with each replicate consisting of a pool of six fully expanded leaves (leaf plastochron index 5–10). tr, treatment; t, time post inoculation; ns, nonsignificant; *, P ≤ 0.05; ***, P ≤ 0.001.