The developmental transition to flowering in Arabidopsis is associated with an increase in leaf chloroplastic...

The developmental transition from vegetative growth to flowering in Arabidopsis is associated with a precipitous decline in the activity of leaf ascorbate peroxidase (APx), an enzymatic scavenger of hydrogen peroxide, and an increase in specific lipid peroxidation leading to the accumulation of 13-hydroperoxy-9,11,15 (Z,E,Z) octadecatrienoic acid (13 HOO-FA). The appearance of this specific isomer suggests that it is of enzymatic origin and may represent the activation of an oxylipin signaling pathway. We thus hypothesized that leaf 13-lipoxygenase (LOX) activity increases at the floral transition and leads to the observed elevation of 13-HOO-FA levels. Leaf protein extracts were prepared from seven distinct life stages of Arabidopsis plants and used to assay for LOX activity. We report that leaf 13-LOX enzymatic activity increases two- to three-fold from the vegetative stage to the immediate post-floral transition stage. We found two forms of LOX activity in cell extracts and show that the higher pH optimum form is the isoenzyme activated. This increase is correlated with a small increase in H(2)O(2), perhaps resulting from the previously reported decline in leaf APx activity. Very low levels of exogenous H(2)O(2) activate the induced form in vegetative leaf extracts in vitro, suggesting that the floral transition-dependent APx decline and subsequent H(2)O(2) elevation are involved in activating plastid 13-LOX and thus a second messenger oxylipin pathway.