Abstract: To regulate the decomposition of organic nitrogen in tobacco soil, 52 strains of ammoniated bacteria were isolated from the soil of Weining and Meitan. After screening by the Nessler's reagent method and the ammoniated culture medium culture method, bacterial strains W2, W4, W6, W12, W16, Z3, Z6, Z15, Z9 and Z50 with strong decomposition effects on organic nitrogen were identified. 16S rDNA sequence analysis indicated that strains W2, W4, W6, W12, W16, Z3, Z6, Z15, Z9 and Z50 belong to Chryseobacterium, Pantoea, Bacillus sq., Bacillus simple, Lysinibacillus, Bacillus sq., Aqrobacterium, Bacillus megaterium, Brevundimonas and Paenarthrobcter. The results from the ammoniated culture medium culture method showed that the organic nitrogen decomposition ability of Z9 was significantly higher (p ≤ 0.05) than that of the other 9 strains after 72 h of culture, with the decomposition effect of 46.1%-93.4%. The results of indoor organic fertilizer culture test showed that the inorganic nitrogen content of the treatment with bacterial strains was higher than that of the treatment without bacteria (CK), the inorganic nitrogen content of the treatment with bacterial strains increased 550.80-1823.71 mg/kg after 65 days of cultivation, and the inorganic nitrogen content increased 1.15-1.49 times by adding bacterial strains. The results of this study can be applied to the configuration of organic nitrogen decomposition composite microbial agents.