Uptake, translocation, and transformation mechanisms of tris(2-chloroethyl) phosphate (TCEP) in hydroponic wheat (Triticum aestivum L.) were systematically investigated using compound-specific stable isotope and multiomics analyses in this study. Results showed that TCEP was quickly adsorbed on root epidermis and then absorbed in roots via water and anion channels as well as an active process dependent on energy. Active process and anion channel preferentially translocated TCEP-containing light carbon isotopes and dominated the transmembrane transport of TCEP to enter vascular bundle. Transcriptomic and metabolomic analyses indicated gene-encoding ATP-binding cassette (ABC) transporters and purple acid phosphatases (PAPs) and glutathione Stransferases (GSTs) involved in TCEP transport and transformation, respectively. Molecular docking simulations showed that TCEP bound to the hydrophilic cavity of ABC transporter/PAP and hydrophobic cavity of GST, and hydrogen bonding was the important driving force. The results of this study offered insights for future effective mitigation of TCEP risk in edible plants.