In order to clarify the translocation and metabolism of glyphosate in lead tree, a mixture of glyphosate (0.5 mmol) and 14C-glyphosate was injected into mini tree plants (1.5-cm trunk diameter and 120 cm tall), simulating a grown lead tree in a greenhouse experiment. Within 6 d after treatment (DAT), all leaves on the lead tree plants dropped. Analysis of nonlabeled glyphosate in both xylem and phloem every 15 d showed that glyphosate residues accumulated mainly in phloem, similar to the findings in established lead tree. In this experiment, glyphosate concentration in the phloem of three parts of the trunk, i.e., upper (15 cm above the upper injection hole), middle (between the two injection holes), and lower (15 cm below the lower injection hole) parts, increased rapidly within 45 DAT, and decreased thereafter. Obviously, significant glyphosate metabolism occurred during the latter period. Further 14C radioactivity measurement also exhibited similar time-course changes of glyphosate dissipation, except that a low level of radioactivity in the phloem of the upper part and a high level of radioactivity in the lower part were detected from 45 to 90 DAT, suggesting that more glyphosate metabolites in the phloem of the upper part might have translocated with photosynthetic assimilates and reallocated to the lower part of the mini tree. Thin-layer chromatography (TLC) analysis of metabolites derived from 14C-glyphosate also revealed that about 70% of the radioactivity recovered in the phloem of the lower part comprised the unknown metabolites M1 (retention factor [Rf] : 0.83) and M2 (Rf : 0.94), with nearly 28% of the radioactivity from sarcosine (Rf : 0.76), and less than 2% of the radioactivity from aminomethylphosphoric acid (AMPA) (Rf : 0.49) and glyphosate (Rf : 0.36). It is concluded that active metabolism of glyphosate in this 90-d simulation experiment occurred mainly from 45 to 90 DAT, and the primary metabolism was modulated by C-P lyase.