Abstract: Continuous cropping obstacle is one of the main restriction factors in potato industry. In order to explore the mechanisms of potato’s continuous cropping obstacle and to reduce the impact  on potato’s tuber yield, a field experiment combined with PCR-DGGE molecular fingerprinting was conducted to investigate the fungal population structure and its biological effect in rhizosphere soil of continuously cropped potato. With the increasing year of potato’s continuous cropping, the numbers of visible bands in rhizosphere fungal DGGE profiles increased obviously. As compared with that of CK (rotation cropping), the operational taxonomic unit (OTU) in treatments of one to five years continuous cropping was increased by 38.5%, 38.5%, 30.8%, 46.2%, and 76.9% respectively, indicating that potato’s continuous cropping caused an obvious increase in the individual numbers of dominant fungal populations in rhizosphere soil. Also with the increasing year of potato’s continuous cropping, the similarity of the fungal population structure among the treatments had a gradual decrease. The sequencing of the fungal DGGE bands showed that with the increasing year of continuous cropping, the numbers of the potato’s rhizosphere soilborne pathogens Fusarium oxysporum and F. solani increased obviously, while the number of Chaetomium globosum, as a biocontrol species, had a marked decrease in the fifth year of continuous cropping. It was suggested that potato’s continuous cropping caused the pathogen fungal populations become the dominant microbial populations in rhizosphere soil, and the rhizosphere microecological environment deteriorated, which in turn affected the root system, making the root vigor and its absorption area reduced, and ultimately, the tuber yield decreased markedly.