Phytolacca americana L. (P. americana) is a manganese (Mn) hyperaccumulator plant discovered in southern China, and knowledge of Mn uptake characteristics and mechanisms on this plant may provide essential and critical information for phytoremediation. Synchrotron radiation X-ray fluorescence spectroscopy (SRXRF) microprobe was empolyed in this study to explore the Mn distribution in the root cross-section of P. americana, and effects of metabolic inhibitors (DNP and Na3VO4) and Ca-channel inhibitor (LaCl3) on Mn uptake of P. americana was also investigated under laboratory conditions. Results showed that P. americana has strong abilities for absorpting and accumulating Mn, and the Mn concentration in root, stem, and leaf of P. americana may reach up to 402, 208, and 601 mg x kg(-1) DW, respectively, even only treated with 5 micromol x L(-1) Mn. The highest Mn content can be found in the vascular bundle of root, and then the epidermis, while the lowest Mn content can be observed in the cortex. The Mn content increased when shifted from cortex to vascular bundle, indicating that there was an active transportation in Mn absorption of P. americana root, and the inhibitory effect of DNP and Na3VO4 on Mn uptake further verified the possibilities of active absorption. The Mn uptake was inhibited by 30% with LaCl3, suggesting that Mn uptake in P. americana also closely related to the Ca-channel.