Milk thistle belonging to the Asteraceae family, is an annual plant and prefers light and fertile soils. In usual agricultural systems, the purpose of using fertilizers and their combination is to obtain the highest seed yield. In order to investigation of the effect of mycorrhiza and nano zinc oxide, this research was carried out in Research Farm of Maragheh University during 2017-2018 growing season as Factorial experiment conducted based on a randomized complete block desing with three replications. The investigated factors in this research include the Control (No application of mycorrhiza and nano zinc oxide), Piriformospera indica + Trichoderma spp, Piriformospera indica and Trichoderma spp and nano zinc oxide. The results obtained from this research showed that seed yield and the number of capitols in sub branches were affected by the inoculation of mycorrhiza and also with the application of zinc at the p < 0.01 level. Also, the harvest index was not affected by any of the factors. Also ascorbate peroxidase and catalase were affected by interaction effect of mycorrhiza × zinc at the p < 0.01 level. According to the results, the highest seed yield and number of capitols in sub branches obtained from Piriformospera indica + Trichoderma spp and with application of zinc. The lowest amount of traits was obtained from the control (No application of mycorrhiza and nano zinc oxide). Also, the highest and lowest activities of ascorbate peroxidase and catalase enzymes were obtained from the application of two fungi with nano zinc oxide and control (No inoculation and Zn application), respectively.