Fractionations of rare earth elements (REEs) in wheat (Triticum aestivum L.) were observed through application of exogenous mixed REEs under hydroponic conditions. Middle REE (MREE), light REE (LREE), and heavy REE (HREE) enrichments were found in roots, stems, and leaves, respectively, accompanied by the tetrad effect (an effect that can cause a split of REE patterns into four consecutive segments) in these organs. Investigations into REE speciation in roots and in the xylem sap with X-ray absorption spectroscopy (XAS) and nanometer-sized TiO2 adsorption techniques, associated with other controlled experiments, demonstrated that REE fractionations in wheat were caused by the combined effects of chemical precipitation, cell wall absorption, and solution complexation by organic ligands in the xylem vessels. REE fractionations in wheat, which were derived from the small differences of chemical properties across REE series, may reflect a sensitive internal chemical environment that influences plant accumulation for REEs and their analogues actinide radionuclides.