The potential of biological soil disinfestation (BSD) in control of continuous potato monoculture barrier was investigated in present study. BSD involves the induction of soil reduction conditions through incorporation of easily decomposed organic materials into soil, flooding the soil by irrigation, and covering the soil surface with plastic film. Control (CK) was left without cover and organic amendment as well as flooding. Field experiment was conducted for testing the effect of BSD approach on the control of continuous potato monoculture barrier, especially on tube yield, plant growth and development, suppression of soil-borne pathogen, and soil microbial community and enzyme activities. Compared with CK, BSD treatment significantly increased tuber yield by 16.1% and plant biomass by 30.8%, respectively. Meanwhile, the incidence of diseased plant and the ratio of diseased tuber in BSD treatment also significantly decreased by 68.0% and 46.7% as compared to those in CK, respectively. BSD treatment significantly increased the content of chlorophyll and branch numbers per main stem of potato plants, improved the morphological characteristics of potato root system. In the course of BSD before potato sowing, soil pH value and bacteria/fungi significantly increased, but populations of fungi and Fusarium sp. significantly decreased compared with CK. There were no significant changes in populations of bacteria and actinomycetes between CK and BSD treatments. During potato growing stage, the populations of both soil fungi and Fusarium sp. were lower in BSD treatment than those of CK. With the advance of potato growth, the population of Fusarium sp. in BSD treatment gradually increased compared with CK. There were no significant changes in soil enzyme activities in the course of BSD before potato sowing and the whole of potato growing stage. It was concluded that BSD has the potential to control continuous potato monoculture barrier and may be an important element in a sustainable and effective management strategy for potato soil-borne diseases.