Rice spikelet rot disease caused by Fusarium proliferatum not only leads to yield reduction, but also contaminates rice grains with mycotoxins. In this study, the effects of exogenous amino acids on the mycelial growth and fumonisins biosynthesis of F. proliferatum were determined through the addition of amino acids to the growth media. The results showed that the growth of F. proliferatum was affected by different exogenous additives and different addition amounts compared to the mock control. The mycelial growth of F. proliferatum was obviously inhibited in MM media supplied with L-arginine, L-glutamate, L-proline and L-citrulline, respectively, and in PDA media supplied with L-glutamate. Fumonisins production was significantly increased by different kinds of amino acids. Fumonisins content tended to reduce in PDA media and increase in MM media with the higher concentration of amino acids. There were significant changes in expression levels of the fumonisins biosynthetic (fum) genes treated with exogenous amino acids, but expression patterns of fum genes were not consistent with fumonisins production. The results demonstrated that the mycelial growth and fumonisins production of F. proliferatum were influenced by the quantity and type of amino acids, and the correlation between different amino acid composition and mycelial growth fumonisins biosynthesis was different.