In order to determine whether hydrogen peroxide (H2O2) generated by polyamine oxidative degradation is involved in the development of lateral roots in soybean, the length and the number of lateral roots, the activities of polyamine oxidases and diamine oxidases, and the endogenous free polyamine and H2O2 content were analyzed in soybean (Glycine max (Linn.) Merr.) main roots of 2-d-old seedlings after treatments for 2 d with exogenous β-hydroxyethylhydrazine (an inhibitor of polyamine oxidases), H2O2, putrescine, cyclohexylamine (an inhibitor of spermidine synthase) or N,N'-dimethylthiourea (a scavenger of hydrogen peroxide).β-hydroxyethylhydrazine treatment strongly inhibited the development of lateral roots in soybean seedlings,reduced the activities of polyamine oxidases and diamine oxidases, decreased H2O2 levels, and led to the accumulation of endogenous polyamines in the main roots. The inhibitory effect of β-hydroxyethylhydrazine on root development could be alleviated by exogenously applied 10 μmol/L H2O2 (a major product of polyamine oxidation). Treatment with cyclohexylamine and putrescine promoted root growth slightly, but treatment with cyclohexylamine plus N,N'-dimethylthiourea or putrescine plus N,N'-dimethylthiourea prevented the development of soybean lateral roots. The effects of these treatments on the development of soybean lateral roots were consistent with the changes in endogenous H2O2 levels. These results suggest that the development of soybean lateral roots is associated with the oxidative degradation of polyamines, and that their products,especially H2O2, are likely to play an important role in the growth of soybean lateral roots.