Brassica carinata CIL1 mediates extracellular ROS production during auxin- and ABA-regulated lateral root development

In the plant cytoplasm signals from multiple sources interact to control development. Lateral root proliferation is controlled by the antagonistic action of the hormones auxin and abscisic acid (ABA). Auxin stimulates the separation of pericycle initials and cell expansion. ABA is required for lateral root initiation and elongation and acts as an antagonist through auxin-dependent pathways. These hormones also mediate reactive oxygen species (ROS) accumulation in growing roots and promote cell expansion. We describe the isolation and characterization of a copper- and auxin-induced gene, COPPER INDUCED in LEAVES (CIL1), from Brassica carinata. Transgenic B. carinata seedlings expressing antisense CIL1 were used to determine the biological function of this gene. Lines with reduced CIL1 expression showed a decrease in lateral root development, as well as reduced sensitivity to auxin and ABA. Steady-state analysis of redox components showed a decrease in NADPH oxidase, superoxide dismutase, and catalase activity, accompanied by an increase in hydrogen peroxide concentration. GFP-tagged CIL1 accumulated at the plasma membrane and in the apoplast indicating CIL1 is likely extracellular. From these data we propose that CIL1 is an extracellular protein involved in ROS cycling mediating auxin and ABA signals.