Fusarium head blight (FHB) epidemics on wheat have been recurrent in subtropical southern Brazil. The contamination of grains by mycotoxins associated with FHB has gained increasing importance in the Brazilian flour market and wheat–derived products. Efforts have been directed towards developing genetic resistance in wheat against FHB and mycotoxin contamination, particularly concerning deoxynivalenol (DON) and zearalenone (ZEA). However, the diversity of Fusarium species associated with FHB in southern Brazil must be considered due to their potential to produce different mycotoxins. A published study involving 317 strains from Paraná State highlighted the predominance of the F. graminearum species complex (FGSC) as FHB pathogens. Fusarium graminearum s. str. of the 15-acetyl(A)DON genotype accounted for 69%, followed by F. meridionale of the nivalenol (NIV) genotype at 19%, F. cortaderiae of the NIV and 3-ADON genotypes at 8%, and F. austroamericanum of the 3-ADON genotype at 3%, all of which are type B trichothecene producers. Other species included F. poae (type A trichothecene), F. tricinctum, F. fujikuroi, and F. incarnatum-equiseti complexes, each comprising less than 1%. This diverse array of species and trichothecene genotypes has influenced the inoculum composition used in genetic resistance trials. This study aimed to determine the range of mycotoxin contamination, including type A and B trichothecenes and ZEA, in wheat grains (whole grain flour) from experimental plots testing cultivar resistance to FHB from 2019 to 2023 in the southern region of Paraná. These plots were artificially inoculated with a macroconidial suspension of 10 isolates, representing the diversity of FGSC species and trichothecene genotypes described above, with concentrations ranging from 0,35–6,7x10^4 spores per mL, applied to wheat at the mid-anthesis stage. Mycotoxin content was measured using standard protocols at the Central Laboratory of Agraria Cooperative with an Acquity UPLC I-Class Plus System. DON was the predominant mycotoxin, with levels varying across genotypes and years [below detection level (BDL)–33341 µg/kg], alongside 15-ADON (BDL–320 µg/kg) and NIV (BDL–2887 µg/kg). Notably, 15-ADON and 3-ADON were detected only in the last three years, while NIV and DON-3-glucoside (D3G) (BDL – 3561 µg/kg) levels showed more stability than DON over the years. Type A trichothecenes such as diacetoxyscirpenol (DAS), T-2, and HT-2 mycotoxins were absent. In conclusion, DON is a primary concern, although NIV, D3G, and ZEA warrant attention, and the relevance of 15-ADON and 3-ADON requires further study. This information will be useful for mycotoxin management in Brazil.