This work aimed to study the role of arbuscular mycorrhizal fungi (AMF) in Hg and major mineral nutrient uptake and tissue localization of these elements in the roots of maize plants. Maize plants were grown in pots filled with non- and Hg-contaminated substrate (50 μg Hg g-1 as HgCl2) and inoculated with two types of AMF inocula: a) Glomus sp. originating from Hg-polluted soil of a former Hg smelting site in Idrija, Slovenia, and b) commercial AM inoculum Symbivit. Controls were inoculated by corresponding bacterial extracts only. Tissue localization of Hg and major mineral nutrients was performed by laser ablation-inductively coupled plasma-mass spectroscopy (LA-ICP-MS) on cryofixed and freeze-dried root cross-sections. AMF colonization increased plant biomass in non-contaminated substrate, while this effect was not seen in Hg-contaminated substrate. Hg increased total plant biomass more than AMF inoculation, possibly through hormetic effects. AMF increased Hg uptake into the roots, as well as Hg transfer to the shoots. AMF affected plant mineral nutrient uptake, depending on the type of AMF inoculum and the presence of Hg. In the roots, Hg was mainly localized in rhizodermis and endodermis, followed by the cortex and the central cylinder. Higher Hg concentrations were detected in the central cylinder of AM plants than in that of the controls, pointing to a higher Hg mobility and potential bioavailability in AMF inoculated plants.