Propyl 4-(2-(4,6-demethoxypyrimidin-2-yloxy)benzylamino) benzoate (ZJ0273), is a novel broad-spectrum herbicidedeveloped for oilseed crop.In this study, fifteen distinct isolates capable of utilizing ZJ0273 as the sole carbon source were isolated from crop soil.One of the isolates, belonged to the family Amycolatopsis and designatedAmycolatopsis sp.M3-1, had higher ZJ0273-degarding ability and was used for the further study.At 30 ℃ and pH 7.0, degradation rate of ZJ0273 by Amycolatopsis sp.M3-1 could reach 59.3% and 68.5 % in 25 day and 60 day, respectively.In order to identify the intermediates and study biodegradation pathway of ZJ0273 by Amycolatopsis sp.M3-1, 14C labeled on different rings, i.e., [4,6-14C-pyrimidine] ZJ0273 and [U-14C-benzyl] ZJ0273, ultralow liquid scintillation counting and LC-MS/MS were used in the biodegrading metabolism experiments in aquatic system.