This study aimed to study the relationship between allelopathic bacteria and Rehmannia glutinosa growth, and clarify the causes of R. glutinosa continuous cropping cbstacle by using soil microbial plate isolation technique and 16S rRNA gene analysis method. From 1 year R. glutinosa replanted soil, 19 strains of bacteria were extracted, two of which were identified to be Pseudomonas putida DH03 and Enterobacter amnigenus DH05. Strain DH03 and strain DH05 didn’t infect R. glutinosa leaves. However, their metabolites had toxic effects on both leaves and tubes of R. glutinosa, and were typical allelopathic bacteria. Meanwhile, constitutes of R. glutinosa had a positive effect on the growth of strain DH03 and strain DH05. Based on the interactive effects between allelopathic bacteria and R. glutinosa, the hypothesis was proposed on the allelopathic bacterial effect which had caused the R. glutinosa continuous cropping cbstacle. Allelopathy of allelopathic bacteria was one of the main factors that had caused R. glutinosa continuous cropping obstacle.