FUM gene expression and variation in fumonisin production of clonal isolates of Fusarium verticillioides MRC 826

B-series fumonisins (FBs) are a family of carcinogenic mycotoxins that commonly occur in maize. These mycotoxins cause multiple diseases in animals and are epidemiologically associated with several human diseases in populations for which maize is a dietary staple. FBs are produced by multiple genera of the fungi Aspergillus, Fusarium and Tolypocladium, but the plant pathogen Fusarium verticillioides is considered the primary cause of FB contamination in maize. One F. verticillioides strain, MRC 826, is reported to produce high levels of FBs. However, in the current study, 18 isolates derived from strain MRC 826 exhibited highly variable levels of FB, which negatively correlated (r=-0.333; P<0.008) with fungal growth. Microsatellite analysis confirmed that all MRC 826 derived isolates examined were clonal, and 100% DNA sequence identity was observed across the FUM gene clusters of two high FB producing and two low FB producing isolates. At the gene expression level, qRT-PCR at each time point (7, 14, 21 and 28 days of incubation) showed differential upregulation of selected FUM genes in the high compared to the low FB isolates. Variation in FB production appears due to differences in FUM gene expression, most likely caused by sequence differences at unexamined loci not part of the FUM cluster or from epigenetic influences. Clarification of the genetic/epigenetic basis for quantitative differences in fumonisin production among strains and isolates of F. verticillioides has potential to reveal targets for reducing FB contamination in maize.

Effects of Muffin Processing on Fumonisins from 14C-Labeled Toxins Produced in Cultured Corn Kernels

The persistence of fumonisins during cooking is known to be affected by several factors, including thermal degradation and the presence of various ingredients in corn-based food recipes that can react with the toxin. A method for the production of corn kernels containing 14C-fumonisins was developed. The corn kernels were colonized by Fusarium verticillioides MRC 826 and supplemented with 1,2-14C-sodium acetate. The specific activity of 14C-FB1 produced made the study of its fate in cornmeal muffins possible. The double-extraction acetonitrile-water-methanol/immunoaffinity column/ophthaldialdehyde high-performance liquid chromatography (HPLC) method was used to determine FB1 levels in cornmeal muffins. Reductions in FB1 levels in muffins spiked with 14C-labeled and unlabeled FB1 (43 and 48%, respectively) were similar, indicating that the extraction method was efficient and consistent with previous reports. However, with the labeled corn kernel material, recovery levels based on the 14C counts for the eluate from an immunoaffinity column were much higher (90%). This finding indicates that some fumonisin-related compounds other than FB1 that were present in the cornmeal were recognized by the antibodies but not by the HPLC method.