Fumonisins and AAL toxins are polyketide-derived mycotoxins produced by several important fungal pathogens of crop plants. The toxins contain a linear, dimethylated polyketide chain, but differ in chain length and methylation positions. Here, we have isolated the major metabolites from a fumonisin null mutant, Fusarium verticillioides strain 5777, that has been transformed with the AAL-toxin polyketide synthase gene (ALT1). The results showed that the metabolites maintained the chain length and methylation pattern of fumonisins. This suggests that the timing and regioselectivity of the methyltransferase of the ALT1 PKS could be reprogrammed in this Fusarium transformant.