Volume 7, Issue 7, Pages 1191-1210 (July 2014) Spatial H2O2 Signaling Specificity: H2O2 from Chloroplasts and Peroxisomes Modulates the Plant Transcriptome Differentially  Sewelam Nasser , Jaspert Nils , Van Der Kelen Katrien , Tognetti Vanesa B. , Schmitz Jessica , Frerigmann Henning , Stahl Elia , Zeier Jürgen , Van Breusegem Frank , Maurino Veronica G.   Molecular Plant  Volume 7, Issue 7, Pages 1191-1210 (July 2014) DOI: 10.1093/mp/ssu070 Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

Figure 1 H2O2 Accumulation and Photoinhibition in GO5 and cat2-2 Plants. (A) DAB staining showing H2O2 production in wild-type (Col-0), GO5 and cat2-2 after transfer of high-CO2-grown plants to normal air at 75 μmol quanta m–2 s–1. (B) Maximum quantum yield of PSII (FV/FM) determined in Col-0, GO5, and cat2-2 at different time points after transfer of high-CO2-grown plants to normal air and of the same plants maintained in high-CO2 conditions. Molecular Plant 2014 7, 1191-1210DOI: (10.1093/mp/ssu070) Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

Figure 2 Comparative Metabolic Profile of Col-0, GO5 and cat2-2 Plants. (A) Heat map of individual metabolites comparing ambient CO2 and high CO2 in wild-type (Col-0), GO5, and cat2-2 plants (log2-fold change). (B) PCA score plot of GC/MS metabolite data. (C) PCA loadings plot of GC–EI–TOF–MS metabolite data. Molecular Plant 2014 7, 1191-1210DOI: (10.1093/mp/ssu070) Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

Figure 3 Representations of the Number of Induced Genes in GO5 and cat2-2 Plants and Their Classification in Different Functional Categories. (A) Venn diagram of the number of H2O2-responsive genes up-regulated only in GO5 or cat2-2 and in both, GO5 and cat2-2, after 8h of H2O2 production. (B) Pie chart of the functional categories of genes whose expression was up-regulated specifically by chloroplastic-produced H2O2. (C) Pie chart of the functional categories of genes whose expression was up-regulated by both chloroplastic- and peroxisomal-produced H2O2. (D) Pie chart of the functional categories of genes whose expression was up-regulated specifically by peroxisomal-produced H2O2. Molecular Plant 2014 7, 1191-1210DOI: (10.1093/mp/ssu070) Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

Figure 4 Dynamics of the Transcriptional Responses of Selected Genes in GO5 and cat2-2 Plants Analyzed by qRT–PCR. The asterisk (*) indicates statistically significant difference calculated by the two-tailed Student’s t-test (P < 0.05) to the time point t = 0. Molecular Plant 2014 7, 1191-1210DOI: (10.1093/mp/ssu070) Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

Figure 5 Schematic Representation of the Camalexin and Indolic Glucosinolate Biosynthetic Pathways Showing the Genes Up-Regulated by Chloroplastic-Produced H2O2. FC indicates the fold-change induction of gene expression with respect to the wild-type. I3M = indol-3-ylmethyl glucosinolate = Glucobrassicin; 4OH-I3M = 4-hydroxy-indol-3-ylmethyl glucosinolate = 4-hydroxyglucobrassicin; 4MO-I3M = 4-methoxy-indol-3-ylmethyl glucosinolate = 4-methoxyglucobrassicin. Molecular Plant 2014 7, 1191-1210DOI: (10.1093/mp/ssu070) Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

Figure 6 Diagram Showing the Spatial H2O2 Signaling Effects Occurring in the GO5 and cat2-2 Plants after 8h of Moderate Production of H2O2. H2O2 originated in chloroplasts or peroxisomes can induce two different kinds of responses: one which is dependent on the subcellular production site and the other which integrates H2O2 signals independently from the subcellular site of its production. Molecular Plant 2014 7, 1191-1210DOI: (10.1093/mp/ssu070) Copyright © 2014 The Authors. All rights reserved. Terms and Conditions