Genes encoding homologs of the gp91phox subunit of the plasma membrane NADPH oxidase complex have been identified in plants and are hypothesized to be a source of reactive oxygen species during defense responses. However, the direct involvement of the gene products in superoxide (O2−) production has yet to be shown. A novel activity gel assay based on protein fractionation in native or sodium dodecyl sulfate (SDS)-denaturing polyacrylamide gels was developed. In native polyacrylamide gel electrophoresis, one or two major O2−-producing formazan bands were detected in tomato (Lycopersicum esculentum Mill. cv Moneymaker) and tobacco (Nicotiana tabacum var. Samsun, NN) plasma membranes, respectively. Denaturing fractionation of tomato and tobacco plasma membrane in SDS-polyacrylamide gel electrophoresis, followed by regeneration of the in-gel activity, revealed NADPH-dependent O2−-producing formazan bands of 106-, 103-, and 80- to 75-kD molecular masses. The SDS and native activity bands were dependent on NADPH and completely inhibited by diphenylene iodonium or CuZn- O2− dismutase, indicating that the formazan precipitates were due to reduction by O2− radicals catalyzed by an NADPH-dependent flavin containing enzyme. The source of the plasma membrane activity bands was confirmed by their cross-reaction with antibody prepared from the C terminus of the tomato gp91phoxhomolog. Membrane extracts as well as the in-gel NADPH oxidase activities were stimulated in the presence of Ca2+. In addition, the relative activity of the gp91phox homolog was enhanced in the plasma membrane of tobacco mosaic virus-infected leaves. Thus, in contrast to the mammalian gp91phox, the plant homolog can produce O2− in the absence of additional cytosolic components and is stimulated directly by Ca2+.
1 August 2013 in Environmental and Experimental Botany
Background Freshwater comprises about a mere 2·5 % of total global water, of which approximately two-thirds is locked into glaciers at the polar ice caps and on mountains. In conjunction with this, in many instances irrigation with freshwater causes an increase in soil salinity due to overirrigation of agricultural land, inefficient water use and poor drainage of unsuitable soils. The problem of salinity was recognized a long time ago and, due to the importance of irrigated agriculture, numerous efforts have been devoted towards improving crop species for better utilization of saline soils and water. Irrigating plants with saline water is a challenge for practitioners and researchers throughout the world. Scope Recruiting wild halophytes with economic potential was suggested several decades ago as a way to reduce the damage caused by salinization of soil and water. A range of cultivation systems for the utilization of halophytes have been developed, for the production of biofuel, purification of saline effluent in constructed wetlands, landscaping, cultivation of gourmet vegetables, and more. This review critically analyses past and present halophyte-based production systems in the context of genetics, physiology, agrotechnical issues and product value. There are still difficulties that need to be overcome, such as direct germination in saline conditions or genotype selection. However, more and more research is being directed not only towards determining salt tolerance of halophytes, but also to the improvement of agricultural traits for long-term progress.
Plant respiratory burst oxidase homologs (Rboh) are homologs of the human neutrophil pathogen-related gp91phox. Antisense technology was employed to ascertain the biological function of Lycopersicon esculentum (tomato) Rboh. Lines with diminished Rboh activity showed a reduced level of reactive oxygen species (ROS) in the leaf, implying a role for Rboh in establishing the cellular redox milieu. Surprisingly, the antisense plants acquired a highly branched phenotype, switched from indeterminate to determinate growth habit, and had fasciated reproductive organs. Wound-induced systemic expression of proteinase inhibitor II was compromised in the antisense lines, indicating that ROS intermediates supplied by Rboh are required for this wound response. Extending these observations by transcriptome analysis revealed ectopic leaf expression of homeotic MADS box genes that are normally expressed only in reproductive organs. In addition, both Rboh-dependent and -independent wound-induced gene induction was detected as well as transcript changes related to redox maintenance. The results provide novel insights into how the steady state cellular level of ROS is controlled and portrays the role of Rboh as a signal transducer of stress and developmental responses.