Abstract Potato blackleg disease (PBD) is one of the most serious potato bacterial diseases. Although the importance of the rhizosphere microbiota for plant growth and health is widely acknowledged, the relationship between the composition of the rhizosphere microbiota and PBD is still unclear. To analyze the relationship between the rhizosphere microbiota composition and PBD, we firstly investigated the potato rhizosphere microbiota composition in the settings of PBD, suspected PBD (SPBD), and health using high-throughput sequencing of 16S rRNA genes. We found that PBD was closely related to the composition of the rhizosphere microbiota. Flavobacterium, Acinetobacter, Dickeya, Sphingobacterium, and Myroides were more abundant in the rhizosphere microbiota of potatoes with PBD than in that of healthy potatoes, while Bacillus, Rhodoplanes, Kaistobacter, Pedobacter, and an unidentified genus from Gaiellaceae were less abundant in the setting of PBD. Dickeya was the probable PBD pathogen in the present study. Our results indicated that enhancing Bacillus may be a way to prevent PBD. These findings provide new insights for the diagnosis and prevention of PBD.