Abstract: The effect of Zn² concentration in submerged culture medium on growth of hyphae of Pleurotus. nebridensis and content of Zn² in hyphae was studied in this paper, in order to discuss zinc-rich ability of hyphae of P. nebrodensis and put forward a reference for zinc-rich submerged culture. The hyphae were incubated for two weeks, with different Zn² concentrations from 0 mg/L to 500 mg/L, on a 25 mg/L increase from each level in submerged medium. Then weighing method was used for measuring the biomass of hyphae, and the content of Zn² in dry hyphae was tested by dithizone colorimetry. The results showed that the biomass of hyphae increased with the concentration augment of Zn² in medium. The maximum of the biomass, 1.146 g, emerged at 325 mg/L Zn² concentration, weighed 5.63 times as much as that of the control, and then decreased little by little with the rise of the Zn² concentration. The content of Zn² in hyphae increased with the increase of the Zn² concentration in medium, its maximum was 26.5056 mg/kg, 6.32 times as high as that of the control. There were extremely significant differences between the treatments and the controls. All these showed that there were important effects of the Zn² concentration in medium on the growth of hyphae and the content of Zn² in hyphae. Therefore, the appropriate Zn² concentration was 325 mg/L in medium for zinc-rich culture to hyphae of P. nebridensis. The content of Zn² in hyphae was 24.79 mg/kg at 325 mg/L Zn² concentration in medium, not the maximum, but the hyphae growth would be inhibited if the Zn² concentration in medium was higher than 325 mg/L.