Ectomycorrhizal fungi, a group of widespread symbiotic fungi with plant, obtain carbon source from trees and improve plant mineral nutrient uptake with their widespread hyphal network. Ectomycorrhizal fungi can be used as inoculants to improve the survival rates of plantation. Saprophytic fungi use the nutrition from the debris of plant or animals, and it is difficult to distinguish the saprophytic and ectomycorrhizal fungi by morphological and anatomic methods. In this research, the differences of stable carbon and nitrogen isotopic compositions of these fungi were analyzed. The results showed that the abundances of ¹³C of were higher than those of ectomycorrhizal fungi and the abundances of ¹⁵N of saprophytic fungi were lower than those of ectomycorrhizal fungi. Such differences of stable carbon and nitrogen isotopic compositions between ectomycorrhizal fungi and saprophytic fungi can be ascribed to their different nutrition sources and ecological functions. These results collectively indicate that stable carbon and nitrogen isotopic compositions are an effective proxy for distinguishing between ectomycorrhizal and saprophytic fungi.