Figure 5
Analysis of 18:3-Glu metabolism in steamed leaves and in LOX deficient plants. (a) 18:3-Glu (0.17 nmoles) was applied to either wounded leaves, wounded-steamed leaves, or unwounded leaves. The 18:3-Glu blank sample denotes an unwounded leaf onto which 18:3-Glu was added after freeze killing the leaf. Leaves were extracted after 5 min of the treatments and 18:3-Glu levels were analyzed by LC-MS/MS. Statistical analysis was performed using a non-parametric Moses extreme test with Bonferroni correction, * indicates P < 0.017, N.S. denotes non-significant differences (n = 3, bars denote ± SE). (b) Leaves of irlox2 and irlox3 plants were wounded, 0.17 nmoles of 18:3-Glu were applied on the wounds, and the samples were harvested after 2 min. After extraction, 18:3-Glu derivatives were analyzed by LC-MS/MS (n = 3, bars denote ± SE). Different letters indicate significant differences (univariate ANOVA 13-oxo-13:2-Glu: F3,3 = 85.4 P < 0.001, 13-OH-18:3-Glu: F3,3 = 25.1 P < 0.001 13-OH-18:3-Glu: F3,3 = 136.1 P < 0.001 followed by a Scheffé post-hoc test, P < 0.05). (c) Turnover of 18:3-Glu on WT and irlox2 plants. Leaves were wounded, 18:3-Glu (0.17 nmoles) were applied onto the wounds, and the samples were harvested after different times. 18:3-Glu levels were analyzed by LC-MS/MS. Initial 18:3-Glu amounts (T0) were set at 100% (n = 4, bars denote ± SE).