Blue Light Exposure Targets NADPH Oxidase to Plasma Membrane and Nucleus in Wheat Coleoptiles

Abstract

Wheat coleoptile tips generate superoxide radical as a part of the phototropic response to blue light, but the source of this free radical generation is not known. We evaluated the presence and involvement of homologs of neutrophil NADPH oxidase (NOX), including gp91phox, p22phox, p67phox, p47phox, and p40phox, in wheat coleoptiles using Western blot analysis and immunofluorescence microscopy. Blue light augmented the expression levels of all these subunits and targeted NOX subunits onto the plasma membrane and to the nucleus. gp91phox, p22phox, p67phox, and p40phox showed entry into the nucleus and exhibited physical closeness with DNA. CuZnSOD was also present in the coleoptile tip, which also showed a blue-light-dependent elevation in expression. Superoxide production and phototropic response were both abrogated by DPIC and staurosporine, indicating their cause-and-effect relationship. We conclude that blue light mediates a phototropic response in wheat coleoptiles through modulation of expression of NOX and SOD as well as the translocation of NOX subunits onto the plasma membrane and nuclear membrane. Thus, this study provides a mechanistic explanation for superoxide production during the photoresponse in wheat coleoptiles.