Abstract: Thirty-seven monospora hybrid strains of two Flammulina velutipes varieties were used as experimental materials, the three enzyme activity indexes（cellulase, hemicellulase, laccase） and five agronomic traits indexes（cap diameter, cap thickness, stalk diameter, stalk length and yield） were comprehensive analyzed by descriptive analysis, principal component analysis and cluster analysis, in order to select excellent Flammulina velutipes germplasm resources.The results showed that the coefficient of variation of each index ranged from 20.93% to 42.75%,and the yield index had the largest coefficient of variation（42.75%）.Through principal component analysis and cluster analysis,a comprehensive evaluation system of Flammulina velutifolia was established,and three monospora hybrid strains No.12,No.32and No.33were selected,which could be used as excellent germplasm resources in the future.