The microbial community composition in wheat rhizosphere was analyzed by detecting colony forming units (CFUs) in agar plates. The total CFUs in rhizosphere were 1.04×109/g soil with 9.0×108/g bacteria, 1.37×108/g actinomyces and 3.6×106/g fungi. The 10 dominant bacteria were isolated from wheat rhizosphere and were grouped into genus Bacillus according to their full length 16S rRNA gene sequences. Although belonging to the same genus, the isolated strains exhibited different sensitivities to oxytetracycline. When a series of the rhizosphere soil was exposed under various concentrations of oxytetracycline, the microbial community structure was highly affected with significant decline of CFUs of bacteria and actinomyces (22.2% and 31.7% at 10 mg/kg antibiotic, respectively). This inhibition was clearly enhanced with the increase exposure dosage of antibiotic and could not be eliminated during 30 d incubation. There was no obvious influence of this treatment on fungi population. Among the four soil enzymes (alkaline phosphatase, acidic phosphatase, dehydrogenase and urease), only alkaline phosphatase was sensitive to oxytetracycline exposure with 41.3% decline of the enzyme activity at 10 mg/kg antibiotic and further decrease of 64.3%-80.8% when the dosage over 30 mg/kg.