The present study is devoted to investigate the effect of high lead contamination on the soil microfungal composition of the urban forest. The investigation was performed for the urban forest soil contaminated by following concentrations of lead: 80, 800, 8,000 and 80,000 ppm. The experiments revealed high resistances of Alternaria and Fusarium fungi to Pb contamination of urban soil. Simultaneously was determined the relatively low tolerance of soil Penicillium microfungi along lead concentration gradient (up 80 to 80,000 ppm). Moreover, the phytotoxic effect of Alternaria microfungi cultivated on high-lead contaminated soil (80–8,000 ppm) was assessed. Obtained results showed the significant impact of lead concentrations (>80 ppm) on the phytotoxicity of Alternaria. Decrease in germination of pea seeds and average root growth was more than 85% in the high lead contaminated soils in comparison to the control sample. © Springer International Publishing AG, part of Springer Nature 2019.