Abstract:[Background] Stellaria dichotoma L. var. lanceolata Bge. is a perennial herb of Caryophyllaceae, with dried roots (Stellariae Radix) as the medicinal part. [Objective] To explore the changes of rhizosphere soil microbial community structure of S. dichotoma var. lanceolata planted for different years in genuine production area and the correlations of microbial community with soil physical and chemical factors and active components in Stellariae Radix. [Methods] Illumina high-throughput sequencing of bacterial 16S rRNA gene and fungal ITS sequences were performed to reveal the microbial community structures in the rhizosphere soil samples of S. dichotoma var. lanceolata planted for 1, 2, 3, and 7 years. The soil properties and the content of total flavonoids and total sterols in Stellariae Radix were determined by conventional methods. [Results] As the planting years of S. dichotoma var. lanceolate increased, the bacteria showed little change in the richness and increased diversity, while the fungi presented increased richness and diversity. The dominant bacteria were Actinobacteriota, Proteobacteria, and Chloroflexi, and the dominant fungi were Ascomycota and Basidiomycota in the rhizosphere soil. The dominant microbial genera varied significantly in the rhizosphere soil samples of S. dichotoma var. lanceolata planted for different years. In addition, the content of total sterols and total flavonoids in 3-year-old Stellariae Radix were significantly higher than those in the samples of other planting years. The correlation analysis showed that pH and alkali-hydrolyzable nitrogen had strong correlations with the microbial community in the rhizosphere soil. Amycolatopsis, Mortierella, Preussia, and Penicillium had significantly positive correlations with the main active components in Stellariae Radix. [Conclusion] The study preliminarily revealed the rhizosphere soil microorganisms related to the active components in Stellariae Radix, which provided a theoretical basis for the high-quality production of this herbal medicine.