Abstract: In this study, thifluzamide, a fungicide used to control sheath blight, was selected as a representative pesticide to investigate the effects of amino acids on rice germination, growth, and the uptake, translocation, distribution, and efficacy of thifluzamide in rice using soil pot and hydroponic experiments. We quantified key physiological parameters, including seed germination rate, seedling biomass (fresh weight), and thifluzamide residues in rice tissues. The results showed that the germination rate of rice seeds treated with glycine, methionine, proline, phenylalanine, and threonine was significantly lower than that of the control. In contrast, the germination rate remained largely unchanged when treated with the other 15 amino acids. We also found that the growth of rice shoots was enhanced when treated with serine, tyrosine, cysteine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, and arginine, with tyrosine showing the most significant improvement. Under both hydroponic and soil cultivation conditions, serine and tyrosine significantly promoted the accumulation of thifluzamide in rice shoots. Notably, tyrosine treatment increased the thifluzamide content in rice shoots by 43.3%, outperforming the serine treatment. Further research showed that tyrosine altered the distribution characteristics of thifluzamide in rice tissues, significantly increasing its proportion in the apoplasts and soluble fractions while reducing its presence in cell walls. This suggests that tyrosine enhances the fluidity of thifluzamide within cells and improves its uptake efficiency. This study highlights that tyrosine seed treatment can significantly enhance the uptake and transport efficiency of thifluzamide in rice, thereby improving its utilization efficiency and efficacy against sheath blight. These findings provide valuable theoretical insights for the future application of amino acids in seed processing technologies for rice production.