Location: Toxicology & Mycotoxin Research
Project Number: 6040-42000-046-010-A
Project Type: Cooperative Agreement
Start Date: Aug 1, 2023
End Date: Jul 31, 2024
Objective:
The primary objective is to conduct an untargeted metabolomics study on the fungal plant pathogen and mycotoxin producer, Fusarium verticillioides, to gain greater understanding of the efficacy of prospective biological control strategies to reduce fumonisin contamination in maize crops.
One of the core competences of the Institute of Bioanalytics and Agro-Metabolomics at the University of Natural Resources and Life Sciences (hereinafter "BOKU") lies in the area of analytical chemistry and metabolomics. The BOKU Core Facility for Bioactive Molecules Screening & Analysis provides services using mass spectrometry, stable isotope assisted approaches, and other analyses as needed. Our collaboration will serve to experientially broaden the core competences of BOKU and the said Core Facility in the area of untargeted metabolomics. Further, BOKU is interested in mycotoxin production by plant pathogenic Fusarium spp. during infection of crop plants. The USDA ARS project is novel and of interest due to the examination of a natural metabolite from another fungus, Sarocladium zeae, that inhibits fumonisin production by F. verticillioides. This other metabolite is pyrrocidine, and a better understanding of the molecular mechanisms underlying the inhibition of fumonisin biosynthesis can help to reduce mycotoxin contamination of food and feed.
Approach:
A comprehensive biological experiment consisting of two different fungal genotypes with four different treatments, sampled at four different time points with five replicates each, provides the base materials for the provided BOKU core facility services. Untargeted metabolomics analysis is carried out by application of a well-established stable isotope assisted approach. A pipetting scheme for successful preparation of the sample set including all necessary controls is provided by BOKU. The biological experiment is carried out in the US and samples are shipped in ready-to-measure form to Austria. LC-HRMS will be used for analysis of ~200 biological samples, controls, and standards. The service also includes comprehensive introduction into data evaluation, so that the generated data can be individually evaluated in a targeted manner by the PhD Student (Lily Lofton). Automated data evaluation in respect to the untargeted metabolomics analysis will be carried out with MetExtract II and performed by Core Facility staff. Lily Lofton will be trained accordingly, so that critical assessment of the obtained results can be carried out independently. This results in the following milestones:
M1. Introductory training.
M2. Experimental samples measured and QC checks passed.
M3: MetExtract II evaluation using LC-HRMS instrumentation (QExactive Orbitrap HF).
M4: Return to USDA ARS the comprehensive dataset for continued analysis by the Toxicology and Mycotoxin Research Unit in Athens, Georgia.
