Participant 8 Assistant Contractor to Participant 7 Michael Tausz, Dieter Grill Karin Herbinger, Klaus Remele, Maria Müller Institute of Plantphysiology, University of Graz, Schubertstraße 51, A-8010 Graz, Austria

Investigations Biochemical defense –Ascorbate system –Glutathione system –Tocopherols –Chloroplast pigments Chromosomal damages –Aberration rates

Experiments  Container exposed young beech trees  Branch cuvette exposures  Mature beech trees sampling 2003

 Container exposed young (3-yrs-old) trees exposed at 2 canopy levels: sun and shade -Pigments and tocopherols June, July, September -Ascorbate and glutathione due to insufficient material only September Leaves: Roots: -Ascorbate, glutathione and tocopherols due to insufficient material only September -Chromosomal aberration analysis September interface to partner 7

 Branch cuvette exposures -Pigments, tocopherols, ascorbate and glutathione September in leave samples included in branch cuvette fumigation systems interface to partner 7

 Mature trees -tocopherols May, June, July, September, Oktober -Chromosomal aberration analysis May, June, July, September, Oktober interface to partner 4 who contributes ascorbate and glutathione Leaves: - Pigments and tocopherols May, June, July, September, Oktober Roots:

Chromosomal aberrations - are approved indicators for genetic damages and - could be defined as changes in the structure of chromosoms

Chromosomal aberration analysis Material: root tips of young and old beech trees We adjusted the method - in connection with the staining procedure (double staining: Schiff reagent, Giemsa, or Schiff reagent and carmine acetic acid) -in connection with the identification of chromosomal aberrations in meta- and anaphases (bridge, sticky chromosomes, vagrant, fragment etc)

Aberration types - sticky chromosomes - anaphase bridges - vagrant chromosomes anaphase metaphase

Problems: The cell walls of most tips were lignified and/or mycorrhized – but: enough root tips for the classification of chromosomal aberrations

violaxanthin neoxanthin zeaxanthin antheraxanthin chlorophyll a  -carotene Pigments: Chromatogram chlorophyll b lutein  -carotene

Total chlorophyll

? Containers removed

 -Carotene

  Containers removed

 -Carotene

Containers removed

Neoxanthin Containers removed

Lutein Containers removed

Xanthophyll cycle Zeaxanthin Antheraxanthin Violaxanthin Light-protection Light-harvesting De-epoxidised Epoxidised Protection from excessive light energy

Violaxanthin + Antheraxanthin + Zeaxanthin

Containers removed

Deepoxidstate

Containers removed

 -Tocopherol

First results on pigments and  -tocopherol show differences between sun and shade crown. Shade leaves exhibited higher pigment content and lower deepoxidate state of xanthophyll cyclus. Only marginal differences were found between ozone treatments. Summerised

Outlook for the next months  Investigations of - pigments and tocopherol (remaining sampling dates) - Ascorbate and gluthatione (in young trees, branch cuvetts) - Chromosomal abberations  statistical evaluation  comparison with the results of other participants and interpretation