Effects of climate change, global trade and technological changes in processing industries cause higher occurrence of Aspergillus flavus and aflatoxin B₁ (AFB₁) in cereal crops. Four Triticum species: common wheat (Triticum aestivum L.), spelt (T. aestivum ssp. spelta L.), Khorasan wheat (Triticum turgidum ssp. turanicum Jakubz.) and hybrid wheat (T. aestivum L.– F1) were examined for their response to A. flavus infection and production of AFB₁. The grains were obtained from control and artificially field inoculated wheat with A. flavus isolates (No. 1 and No. 2) in the 2016 vegetation season in the region of Vojvodina (Northern province of Serbia). Spelt wheat showed the strongest response to the infection in comparison to other analysed wheat species due to specific physico-chemical characteristics of the hull. The weakest response to A. flavus infections was noted in Khorasan wheat. The highest AFB₁ level (256 μg/kg) was observed in the dehulled spelt grains, in comparison to other species where the AFB₁ in dehulled grains was not detected. The levels of AFB₁ in spelt were about three times higher in hulls (648 and 97.3 μg/kg, respectively) in comparison to grains (256 and 30.7 μg/kg, respectively) in two inoculation treatments (A. flavus No. 1 and No. 2, respectively). In order to investigate the impact of wheat hulls on development of A. flavus, including the biosynthesis of toxic fungal metabolites, physico-chemical and structural properties of different Triticum spp. hulls were characterised. The highest value of the water absorption index and total dietary fibre were observed in spelt hulls in comparison to other wheat species. Additionally, the height value distribution of the fossilized stomatal apparatus of hulls indicates the diversity of spelt wheat compared to other wheat species.