title: Large-scale protein analysis of European beech trees following four vegetation periods of twice ambient ozone exposure.
abstract: In the present study, we performed a large-scale protein analysis based on 2-DE DIGE to examine the effects of ozone on the leaves of juvenile European beech (Fagus sylvatica L.), one of the most important deciduous tree species in Central Europe. To this end, beech trees were grown under field conditions and subjected to ambient and twice ambient ozone concentrations during the vegetation periods of four consecutive years. The twice ambient ozone concentration altered the abundance of 237 protein spots, which showed relative ratios higher than 30% compared to the ambient control trees. A total of 74 protein spots were subjected to mass spectrometry identification (LC-MS/MS), followed by homology-driven searches. The differentially expressed proteins participate in key biological processes including the Calvin cycle and photosynthesis, carbon metabolism, defense- and stress-related responses, detoxification mechanisms, protein folding and degradation, and mechanisms involved in senescence. The ozone-induced responses provide evidence of a changing carbon metabolism and counteraction against increased levels of reactive oxygen species.
  
  
  CONCLUSIONS
  This study provides useful information on how European beech, an economically and ecologically important tree species, reacts on the molecular level to increased ozone concentrations expected in the near future. The main emphasis in the present study was placed on identifying differentially abundant proteins after long-term ozone exposure under climatically realistic settings, rather than short-term responses or reactions under laboratory conditions. Additionally, using nursery-grown beech trees, we took into account the natural genotypic variation of this species. As such, the results presented here provide information on molecular responses to ozone in an experimental plant system at very close to natural conditions. Furthermore, this proteomic approach was supported by previous studies on the present experiment. Ultimately, the combination of this proteomic approach with several approaches including transcriptomics, analysis of non-structural carbohydrates, and morphological effects contributes to a more global picture of how beech trees react under increased ozone concentrations.
authors.affiliation: Technische Universität München, Section of Forest Genetics, Hans-Carl-von-Carlowitz-Platz 2, 85354 Freising, Germany; Helmholtz Zentrum München, Institute of Biochemical Plant Pathology, Ingolstädter Landstraße 1, 85764 Neuherberg, Germany; Helmholtz Zentrum München, Institute of Soil Ecology, Ingolstädter Landstraße 1, 85764 Neuherberg, Germany. Electronic address: renekerner@mytum.de.
authors.forename: René
authors.initials: R
authors.lastname: Kerner
authors.affiliation: University Children's Hospital (UKBB) at University of Basel, Spitalstrasse 33, 4031 Basel, Switzerland; Endothelial Cell Biology Unit, School of Molecular & Cellular Biology, LIGHT Laboratories, and Department of Applied Mathematics, School of Mathematics, University of Leeds, Leeds LS2 9JT, United Kingdom.
authors.forename: Edgar
authors.initials: E
authors.lastname: Delgado-Eckert
authors.affiliation: Helmholtz Zentrum München, Institute of Biochemical Plant Pathology, Ingolstädter Landstraße 1, 85764 Neuherberg, Germany.
authors.forename: Dieter
authors.initials: D
authors.lastname: Ernst
authors.affiliation: Université de Bordeaux, Centre de Génomique Fonctionnelle, Plateforme Protéome Bordeaux, 33000 Bordeaux, France.
authors.forename: Jean-William
authors.initials: JW
authors.lastname: Dupuy
authors.affiliation: Technische Universität München, Ecophysiology of Plants, Department of Ecology and Ecosystem Sciences, Hans-Carl-von-Carlowitz-Platz 2, 85354 Freising, Germany.
authors.forename: Thorsten E E
authors.initials: TE
authors.lastname: Grams
authors.affiliation: Helmholtz Zentrum München, Institute of Biochemical Plant Pathology Research Unit Environmental Simulation, Ingolstädter Landstraße 1, 85764 Neuherberg, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Barbro Winkler
authors.affiliation: Helmholtz Zentrum München, Institute of Biochemical Plant Pathology, Ingolstädter Landstraße 1, 85764 Neuherberg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Lindermayr
authors.affiliation: Technische Universität München, Section of Forest Genetics, Hans-Carl-von-Carlowitz-Platz 2, 85354 Freising, Germany.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Müller-Starck
mesh_terms: D005060:Europe; D029964:Fagus / Q000378:metabolism*; D013058:Mass Spectrometry; D010083:Oxidants, Photochemical / Q000494:pharmacology*; D010126:Ozone / Q000494:pharmacology*; D018515:Plant Leaves / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics
chemical_list: D010083:Oxidants, Photochemical; D010940:Plant Proteins; D020543:Proteome; D010126:Ozone
keywords: 2-DE DIGE; Proteomics; Systems biology; Tropospheric ozone (O(3)); cby