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Ozone-induced gene expression occurs via ethylene-dependent and -independent signalling

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Abstract

Recent studies suggest that ethylene is involved in signalling ozone-induced gene expression. We show here that application of ozone increased glucuronidase (GUS) expression of chimeric reporter genes regulated by the promoters of the tobacco class I β-1,3-glucanases (GLB and Gln2) and the grapevine resveratrol synthase (Vst1) genes in transgenic tobacco leaves. 5′-deletion analysis of the class I β-1,3-glucanase promoter revealed that ozone-induced gene regulation is mainly mediated by the distal enhancer region containing the positively acting ethylene-responsive element (ERE). In addition, application of 1-methylcyclopropene (1-MCP), an inhibitor of ethylene action, blocked ozone-induced class I β-1,3-glucanase promoter activity. Enhancer activity and ethylene-responsiveness depended on the integrity of the GCC boxes, cis-acting elements present in the ERE of the class I β-1,3-glucanase and the basic-type pathogenesis-related PR-1 protein (PRB-1b) gene promoters. The minimal PRB-1b promoter containing only the ERE with intact GCC boxes, was sufficient to confer 10-fold ozone inducibility to a GUS-reporter gene, while a substitution mutation in the GCC box abolished ozone responsiveness. The ERE region of the class I β-1,3-glucanase promoter containing two intact GCC boxes confered strong ozone inducibility to a minimal cauliflower mosaic virus (CaMV) 35S RNA promoter, whereas two single-base substitution in the GCC boxes resulted in a complete loss of ozone inducibility. Taken together, these data strongly suggest that ethylene is signalling ozone-induced expression of class I β-1,3-glucanase and PRB-1b genes. Promoter analysis of the stilbene synthase Vst1 gene unravelled different regions for ozone and ethylene-responsiveness. Application of 1-MCP blocked ethylene-induced Vst1 induction, but ozone induction was not affected. This shows that ozone-induced gene expression occurs via at least two different signalling mechanisms and suggests an additional ethylene independent signalling pathway for ozone-induced expression of genes involved in phytoalexin biosynthesis.

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Authors and Affiliations

  1. GSF-National Research Center for Environment and Health, Institute of Biochemical Plant Pathology, 85764, Neuherberg, Germany

    Bernhard Grimmig, Maria N. Gonzalez-Perez, Bernd Heidenreich, Christian Langebartels, Dieter Ernst & Heinrich Sandermann Jr.

  2. Bayer Crop Science, Target Research, Alfred-Nobel-Strasse 50, 40789, Monheim, Germany

    Bernhard Grimmig & Rüdiger Hain

  3. CREAF, Facultat di Ciencies, Universitat Autonoma, 08193, Barcelona, Spain

    Maria N. Gonzalez-Perez & Josep Penuelas

  4. Friedrich Miescher Institute for Biomedical Research, a branch of the Novartis Research Foundation, Maulbeerstrasse 33, 4058, Basel, Switzerland

    Gerhard Leubner-Metzger, Regina Vögeli-Lange & Fred Meins Jr.

  5. Institut für Biologie II, Botanik, Universität Freiburg, Schänzlestrasse 1, 79104, Freiburg, Germany

    Gerhard Leubner-Metzger

  6. Syngenta Crop Protection AG, Schwarzwaldallee 215, 4058, Basel, Switzerland

    Regina Vögeli-Lange

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  1. Bernhard Grimmig
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Grimmig, B., Gonzalez-Perez, M.N., Leubner-Metzger, G. et al. Ozone-induced gene expression occurs via ethylene-dependent and -independent signalling. Plant Mol Biol 51, 599–607 (2003). https://doi.org/10.1023/A:1022385104386

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