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University of Natural Resources and Life Sciences, Australia
Title: Automated annotation of the wheat metabolome during Fusarium infection using stable isotopic labeling and custom-tailored data processing workflows

Christoph Bueschl is a Post-doctoral Researcher at Metabolomics group at IFA-Tulln with expertise in automated data processing of LC-HRMS data. He is specialized in data evaluation and software development of stable isotope assisted and LC-HRMS based untargeted metabolomics experiments as well as stable isotope assisted tracer experiments that probe the secondary metabolism of either endogenous or exogenous secondary metabolites in biological systems. His developed software tools are actively being used, steadily improved and extended with new functionality and applied in various projects and co-operations. Besides data processing of LC-HRMS data, one of his research interests is statistical evaluation of large datasets especially in the area of metabolomics research.


Fungi of the genus Fusarium infect crop plants thereby causing plant diseases and contamination of food with its toxic secondary metabolites so-called mycotoxins. To characterize the plant’s metabolic defense mechanisms to Fusarium infection, we have developed several analytical protocols accompanied by custom-designed automated data processing tools. The first workflow, entitled all extract, is designed for the detection of all LC-HRMS accessible metabolites of wheat. It uses either uniformly 13C-labeled or 15N-labeled wheat reference material, which has been grown in-house and has an isotopic enrichment of ~99%, for metabolite detection and annotation (~1.000 carbon- and ~300 nitrogen-containing metabolites). Moreover, the 13C-labeling step enables metabolome-wide internal standardization thereby improving relative quantification and subsequent statistical comparison of the experimental groups. A second workflow, named TracExtract, allows probing the metabolism of exogenous or endogenous 13C-labeled tracer compounds in wheat plants and reports only biotransformation products that the plant has produced from the respective tracer. Using this approach, uniformly 13C-labeled phenylalanine and tryptophan tracers have been used to annotate respective tracer-derived wheat biotransformation products (120 and 60 respectively). This annotation is especially helpful since many of the already known defense-related metabolites in wheat are descendants of these two tracer compounds. Furthermore, the tracer approach was successfully used to investigate the plant’s detoxification mechanisms of Fusarium graminearum most potent mycotoxin deoxynivalenol. A total of nine mostly novel detoxification products were detected. Finally, all information about the wheat metabolites is aggregated thus describing each detected metabolite with its total number of carbon and nitrogen atoms and if it is derived from phenylalanine or tryptophan. This untargeted annotation is an invaluable resource for further investigation of wheat-Fusarium interaction on a metabolic level and enables a more focused investigation of potential novel defense-related metabolites.

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