Abstract: 【Objective】To comprehensively evaluate the agronomic traits and genetic diversity of Stropharia rugosoannulata strains, and screen out high-quality strains suitable for cultivation in Sanming, Fujian, which could provide reference for S. rugosoannulata breeding and its development and utilization. 【Method】The genetic diversity of 17 strains of S. rugosoannulata was analyzed by ITS sequence analysis, antagonistic test and SRAP molecular marker. The agronomic traits such as fruiting body length, pileus diameter, pileus thickness, diameter of upper stipe, diameter of lower stipe and single weight were measured respectively. The agronomic traits of S. rugosoannulata were analyzed by sampling correlation analysis and principal component analysis. 【Result】ITS sequence analysis confirmed that 17 strains were all S. rugosoannulata, which belonged to 3 branches in the evolutionary relationship. The results of antagonistic test showed that there were 87 groups of culture combinations with antagonistic reaction and 49 groups of culture combinations without antagonistic reaction, that was, there were great differences among S. rugosoannulata strains. The genetic similarity coefficient of 17 S. rugosoannulata strains was 0.66-0.92, and the genetic background was rich. When the genetic similarity coefficient was 0.79, the 17 strains of S. rugosoannulata could be divided into 6 categories（I-VI）. The genetic diversity index（H’） of different agronomic traits was greater than 1.50. Among them, the coefficient of variation of single weight was the largest（47.82%）, and the coefficient of variation of single yield was small（14.29%）, indicating that the agronomic traits of S. rugosoannulata had abundant variation and high genetic diversity. Correlation analysis showed that the mycelium morphology was significantly and negatively correlated with the mycelium growth rate in cultivation plates（r=-0.565）（P＜0.05, the same below）, and mycelium morphology was extremely significantly and positively correlated with the mycelium morphology and yield in cultivation bags（r=0.772 and r=0.664）（P＜0.01, the same below）. There was significant negative correlation between mycelium growth rate and yield in cultivation plate（r=-0.524）. There was extremely significant positive correlation between mycelium morphology and yield in cultivation bag（r=0.721）. The results of principal component analysis showed that the contribution rate of the first principal component（PC1） was 50.768 %, and the agronomic traits with higher eigenvectors were fruiting body length and yield. The contribution rate of the second principal component（PC2） was 38.729 %, and the agronomic traits with higher eigenvectors were pileus diameter and the diameter of upper stipe. 【Conclusion】The mycelium of S. rugosoannulata strain D-3 grow uniformly and densely, with fast growth rate, high and stable yield, thick fruiting body stipe, and good overall commodity. It can be used as a parent for the breeding of S. rugosoannulata varieties and is suitable for cultivation and promotion in Sanming, Fujian.