To analyze genetic diversity in 10 species of Ranunculaceae. The genetic diversity and genetic structure of 10 species of Ranunculaceae in 22 populations in Luoyang and surrounding areas were analyzed using primers selected by ISSR molecular markers. The 12 selected primers amplified a total of 116 clear bands, and the proportion of polymorphic bands was 98.1%. The average polymorphism information content (PIC) of the primers was 0.9478. The results of genetic diversity analysis showed that the Shannon information index (I) of 22 populations of Ranunculaceae plants was 0.4367±0.1904, and Nei’s genetic diversity index (H) was 0.2807±0.1481. The above results showed rich polymorphism in all 12 primers, and very rich genetic diversity in the 10 species of Ranunculaceae from 22 populations. The gene flow Nm was 0.3096 and genetic differentiation index Gst was 0.5997, indicating that genetic differentiation mainly derived from diversity within populations, with less gene communication among populations. The Mantel test showed positive correlation between genetic distance and geographical distance (r = 0.2530, P < 0.01). Cluster analysis, principle coordination analysis (PCoA) and population cluster analysis yielded broadly consistent clustering results showing that individuals of the same germplasm were closely related, tending to be clustered into one group first; the second grouping was arranged according to the geographical distance. The genetic diversity of 10 species of Ranunculaceae in 22 populations is very rich. The variation among 22 populations is large, which indicates that the 10 species of Ranunculaceae have a strong ability to adapt to the environment. The combination of the three methods can improve the accuracy of cluster analysis of wild Ranunculaceae samples. This study lays the foundation for rational utilization and resource management of Ranunculaceae.