title: Arabidopsis thaliana deficient in two chloroplast ascorbate peroxidases shows accelerated light-induced necrosis when levels of cellular ascorbate are low.
abstract: Arabidopsis chloroplasts have a multi-layered defense against hydrogen peroxide (H(2)O(2)) that includes a stromal and thylakoid ascorbate peroxidase (sAPX and tAPX). Single and double null mutants in SAPX and TAPX (sapx and tapx) were each crossed with ascorbate deficient vtc2. The single, double and triple mutants did not show visual light stress phenotypes when grown at control or high light intensities (CL and HL; 120 and 1,000 micromol photons m(-2) s(-1)). Upon shift from CL to HL, mesophyll of expanded leaves of the triple mutant bleached within hours, with exclusion of the major vein areas; this contrasts to reported patterns of cell death under ozone treatment and calatase deficiency. tapx-vtc2 and sapx-vtc2, but not tapx-sapx or single mutants, showed limited bleaching. Bleaching and necrosis were accompanied by accumulation of H(2)O(2). Cellular concentrations of alpha-tocopherol, ascorbate and glutathione showed dramatic increase in response to HL in all eight genotypes and the four vtc2 genotypes accumulated more glutathione under CL than the others. Transcript analysis of other ROS responsive genes in vtc2 and the triple mutant showed up to 20-fold induction after transition to HL, generally irrespective of genotype. We conclude that chloroplast APX proteins in Arabidopsis can be effectively compensated by other endogenous H(2)O(2) detoxification systems, but that low cellular ascorbate levels in absence of chloroplast APX activity are detrimental to the cell during excess light.
authors.affiliation: Department of Plant Biology, Cornell University, Emerson Hall 332, Tower Road, Ithaca, NY 14853, USA.
authors.forename: Lisa
authors.initials: L
authors.lastname: Giacomelli
authors.forename: Antonio
authors.initials: A
authors.lastname: Masi
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Ripoll
authors.forename: Mi Ja
authors.initials: MJ
authors.lastname: Lee
authors.forename: Klaas J
authors.initials: KJ
authors.lastname: van Wijk
mesh_terms: D017360:Arabidopsis / Q000201:enzymology / Q000502:physiology / Q000528:radiation effects*; D029681:Arabidopsis Proteins / Q000235:genetics*; D060387:Ascorbate Peroxidases; D001205:Ascorbic Acid / Q000378:metabolism*; D002736:Chloroplasts / Q000201:enzymology*; D018506:Gene Expression Regulation, Plant; D006861:Hydrogen Peroxide / Q000378:metabolism; D008027:Light; D009154:Mutation; D009336:Necrosis; D010544:Peroxidases / Q000235:genetics*; D018087:Plastids / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015854:Up-Regulation