The present work deals with the capability for producing fumonisin by Fusarium proliferatum strains isolated from asparagus in China. Fifty of F. proliferatum strains were randomly selected and incubated on cultures of maize grain and asparagus spear, respectively. Fumonisin levels (FB1 and FB2) were determined by high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). The results showed that all 50 strains produced fumonisins in maize culture within a wide range of concentrations, 10–11,499 µg/g and 2–6,598 µg/g for FB1 and FB2, respectively. On culture of asparagus spear,48 strains (96%) produced fumonisins in the range 0.2–781.6 µg/g and no detected to 40.3 µg/g for FB1 and FB2, respectively. All of F. proliferatum strains produced much higher levels of FB1, FB2 and total fumonisins (FB1 + FB2) in maize grain culture than in asparagus spear culture. Meanwhile, fumonisin B3 (FB3) was identified in all maize culture extracts and most of asparagus spear culture extracts. This is the first study carried out the fumonisin-producing ability of F. proliferatum strains isolated from asparagus in China. The information obtained is useful for assessing the risk of fumonisins contamination in asparagus spear.