The rhizosphere bacterium ZKB-2 showed strong nematostatic activity against Meloidogyne incognita. Our study aimed to identify the nematostatic metabolites and evaluate the biocontrol efficiency in pot experiments. As the bacterial culture filtrate showed 100% nematostatic activity against M. incognita juveniles in 12 hr, we isolated and identified six compounds following activity guiding. 3-methoxycyclobutane-1, 2-dione showed 58.9% and 72.2% nematostatic activities against juveniles of M. incognita in 12 and 48 hr, with strong LC50 value at 447 μg mL-1. In pot experiments, treatments with the bacterial culture filtrate of strain ZKB-2 showed significant efficacy, especially at doses of 150 mL/pot, which were close to that of avermectin (positive control) at 0.01 g kg-1 soil. The most effective treatment inhibited 85.1% population of juveniles of M. incognita in the roots and 76.9% in the rhizosphere soil after 30 days. Furthermore, the promoting tomato growth also significantly increased in a dose-dependent manner. Our results revealed the potential of strain ZKB-2 to act as a biocontrol agent in the integrated management of root-knot nematodes on tomatoes.