【Objective】 Owing to the growth characteristics of wolfberry ( Lycium barbarum L.) per se and the limitation of soil resources in the traditional wolfberry plantation regions, long-term monocropping of the plants has become one of the main factors hindering sustainable development of the industry of wolfberry production and causing serious economic loss and potential eco-problems. Previous studies found that monocropping could significantly affect diversity and composition of the bacterial communities in the rhizosphere of replanted L. barbarum, however, they did not deal much with responses of the fungal communities therein to the long-term monocropping. 【Method】Soil samples were collected from the topsoil layer (0~20 cm) near the plants in the long-term monocropping wolfberry fields and randomly in the control (no wolfberry cultivation history) with a soil auger (n). In order to have enough rhizosphere soils, root segments from at least ten individual plants were collected. Rhizosphere soil adhered to the roots were collected by washing with 10 mmol•L^-1 NaCl solution and centrifuging. The genomic DNA was extracted from 0.1 g wet rhizosphere and bulk soil sample using the PowerSoil DNA isolation Kit, separately. The ITS2 region was amplified and sequenced with the Illumina HiSeq technique. Then metrics of α diversity (Shannon index and OTU richness) and weighted and unweighted UniFrac distances were calculated with the aid of QIIME. And real time PCR was used to quantify abundance of the bacterial and fungal populations in each DNA samples.【Result】 Real time PCR found that monocropping significantly increased abundances of the bacterial and fungal communities in both bulk soil and rhizosphere of L. barbarum as compared with what was found in the control fields (P < 0.05). However, the effect was much higher on fungi than on bacteria, and hence disturbed the balance between bacteria and fungi in ratio, thus making the microbial environments in the rhizosphere and bulk soil tend to be fungal dominant. Sequencing analysis discovered that Ascomycota, Basidiomycota, Zygomycota, Chytridiomycota, and Glomeromycota were the dominant phyla in all the samples, and that the relative abundance of Zygomycota in the rhizosphere of replanted wolfberry was significantly lower than that in the control, whereas an opposite trend was found with Glomeromycota (P < 0.05). And FUNGuild predication of fungal functions demonstrated that the relative of arbuscular mycorrhizal fungi (AMF) in the rhizosphere of replanted wolfberry in the monocropping fields decreased significantly relative to what in the control fields (P < 0.05). Non-metric multidimensional scaling analysis (NMDS) revealed that the fungal communities in bulk soil and rhizosphere of replanted wolfberry were quite similar to each other in the monocropping fields, but not so in the control, although they did not very much in diversity from those in the control. Furthermore, distance based redundancy analysis indicated that soil pH, electrical conductivity, and contents of nitrate nitrogen and readily available phosphorous were the main factors affecting soil fungal communities in the rhizosphere of wolfberry, while contents of nitrate nitrogen and readily available phosphorous were the ones explaining changes in soil fungal communities in the bulk soils (P < 0.05).【Conclusion】All the findings in this study suggest that long-term fertilization in the monocropping wolfberry fields might be one of the major causes driving the succession of soil fungal communities in the rhizosphere of replanted wolfberry and altering the interaction between wolfberry and soil fungal community. Therefore, the study may shed some new light on mechanism of the forming of the hazard of long-term monocropping of wolfberry to sustainable development of the industry of wolfberry production.