Metabolomics

Based on the technical advancement of our cutting edge metabolomics workflows and the outcome of biological studies, we will pursue the following topics: The stable isotope labeling (SIL) assisted LC-HRMS workflows shall be extended to more polar metabolites (HILIC LCHRMS) and a novel MetExtract software module to support the recognition of nonpredictable isotope patterns with a lower degree of enrichment. In cooperation with F3705/LAP3714 and F3711 we will intensify the integrated evaluation of transcriptomics and metabolomics data to elucidate associated metabolic processes and derive putative novel pathways. Together with F3708 and F3711/Lemmens, the phytolabelbox will be used for the custom-tailored production of globally ¹³C enriched plant material for maximum metabolome coverage. Based on the finding that Fusarium can hydrolyze phenylpropanoid conjugates such as coumaroyl-tryptamine and efficiently transform tryptamine to indole-3-acetic acid, the metabolism of U-¹³C tryptamine / U-¹³C indolacetic acid will be investigated and the role of auxins in FHB shall be studied together with F3702, F3708 & F3711. Both the metabolism of the ¹³C / ¹⁵N labeled plant endogenous defense compounds phenylalanine and putrescine as well as the synergism of ¹³C labeled fungal secondary metabolites (i.e. DON with culmorin and ZEN) and their effect on the plant metabolome shall also be studied in cell cultures and flowering wheat using our established SIL assisted metabolomics workflows. The metabolism of DON, culmorin, butenolide and ZEN in mammalian cell cultures will be investigated in cooperation with F3718. Moreover, together with F3702 we also intend to use our isotope assisted workflow to elucidate polyketide-type secondary metabolites of F. graminearum, putatively involved in FHB pathogenesis. Stable isotope assisted metabolite profiling of double knockout mutants, deficient in H3K27 methyltransferase and selected, infection associated polyketide synthases (provided by F3702) should lead to the identification of currently unknown, tentatively bioactive fungal metabolites. The LC-MS and GC-MS based analytics in support of project partners shall be continued in the next funding period.