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Working Plans & Goals Project year 2004 Metabolic profiling of Arabidopsis thaliana :: post-doc Jens Rohloff AIMS OF RESEARCH – Model plant Arabidopsis thaliana C haracterize secondary metabolites in At with focus on the glucosinolate-myrosinase complex I nvestigate T-DNA-knockout mutants being studied within The Plant Genetics Group at NTNU to sustain the group’s molecularbiological work S tudy both abiotic and biotic factors affecting plant growth and metabolism regarding morphology and ontogenetic development
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Metabolic profiling of Arabidopsis thaliana :: post-doc Jens Rohloff The glucosinolate-myrosinase complex Glucosinolates derived from amino acids - leucine - methionine - phenylalanine - tryptophane Altered pathway of GLS induces changes in biosynthesis of other secondary metabolites involved in - plant defense - plant-insect interactions - stress response Detection of concentration levels of - Glucosinolates - Plant hormones - Amino acids - Volatiles, i.e. isothiocyanates, nitriles, terpenes, hydrocarbons indole glucosinolates
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Metabolic profiling of Arabidopsis thaliana :: post-doc Jens Rohloff Preliminary Results 2003 - Volatiles from Arabidopsis Metabolic Profiles from A.Arabidopsis mutants (wt,TGG1, TGG2,Cyp83A1,Cyp83B1) B. Ecotypes of Arabidopsis (Col, Cve, Ler, Was) C. Plant organs (leaves, stems and flowers) D. Living plants (in vivo) or Autolysed plants Identified Volatiles Terpenes Green leaf volatiles Isothiocyanates + Nitriles Hydrocarbons (alkanes, aldehydes ketones,esters)
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Metabolic profiling of Arabidopsis thaliana :: post-doc Jens Rohloff Gas chromatography analysis (GC-MS) of volatiles from Arabidopsis mutant Cyp83A1 Method - headspace solid-phase microextraction Z-3-hexenol Z-3-hexenyl acetate 4-methyl pentyl ITC 2-ethyl benzyl ITC alpha-phellandrene para-cymene hexyl ITC heptyl ITC 3-methyl butyl ITC nonyl ITC 1-hexanol... over 100 chemical compounds have been tentatively identified by mass spectral database search (GC-MS) ! decanal nonanal tetradecanal
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Co-operation within BIOEMIT and outside the project 1. The Plant Genetics Group, Dep Biol, NTNU Ishita Ahuja (res.) + Anna Kusnierczyk (dr.stip.) plant-insect interactions Anna Vera De Felice (post-doc) metabolomics 2. Institute of Physical Chemistry, Dep Chemistry, NTNU Prof. Bjørn Alsberg, Endre Anderssen (post-doc) bioinformatics 3. The Plant Biocentre, Dep Biol, NTNU Prof. Tor-Henning Iversen, Knut Robert Fossum (res.) space biology 4. Neuroscience Lab, MTFS, NTNU Stig Ulland (dr.stip.) plant-insect interactions 5. Group of Ecological Chemistry, Dep Org Chemistry, KTH Prof. Anna-Karin Borg Karlson biochemistry Metabolic profiling of Arabidopsis thaliana :: post-doc Jens Rohloff
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First publication title (submission to Phytochemistry until April 2004) “Altered headspace volatile profiles of Arabidopsis mutants with deficiencies in the glucosinolate biosynthetic pathway” Conference 15. Int Conf Arabidopsis Research, July 10 - 14, 2004, Berlin Fellowship abroad (3 up to 6 months in 2005) Max-Planck-Institute at Jena or Potsdam, Germany >>> funding will be applied via EU (Marie Curie), NFR or DAAD
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