Combining with seed coating method, rhodamine B (RB), a kind of fluorescent compounds had beenused in seed labeling as an anti-counterfeiting technology. Effects of broad bean seed coating with RB onphysiological parameters and fluorescence performance in broad bean seedlings were performed. Theprolonged fluorescence in vascular bundles of seedlings treated with RB could be used as a marker ofseed anti-counterfeiting. The experiment showed that RB had no negative effect on seed germination,seedling growth, seedling protective enzyme activities, and the contents of malondialdehyde (MDA) andchlorophyll, after coating seeds with ratio 1 kg RB to 10–30 kg seeds. In addition, the RB adsorption bysoil was accorded with the isothermal adsorption equations of Langmuir, which exhibited the linearcorrelation as 0.9959, the maximum adsorption as 1.716 mg/g, and the adsorption equilibrium constantas 0.042 L/mg. The soil adsorption of RB could be effectively degraded by UV/Fenton. Present researchsuggested that coating broad bean seeds with appropriate concentration RB were safe to seed germinationand this technology was suitable for anti-counterfeiting application.