New and improved strategies are needed to detect plant pathogens before symptom onset. The fungus Fusarium graminearum produces trichothecene mycotoxins that pose a threat to the health of domestic animals and humans. This research aims to address knowledge gaps pertaining to early detection of pathogens in agricultural settings and the feasibility of using callus culture as a conduit for pathogen detection and mycotoxin characterization. Calli and whole plants of the Landsberg erecta accession of Arabidopsis thaliana were infected with two strains of F. graminearum (3-ADON and 15-ADON genotypes), and mycotoxins were quantified using Gas Chromatography Mass Spectrometry (GC-MS). It is expected that concentrations will be congruent between whole plants and calli, with the greatest concentration of trichothecene mycotoxins being produced in the 15-ADON strain due to the greater prevalence in this genotype. If successful, our work has the potential to demonstrate that callus culture is a viable approach to studying the pathosystem in plants, paving the way for a faster method for estimating F. graminearum trichothecene production in planta.