Identification of incorrect gene models in the F. graminearum database. A. Statistical analysis of the incorrect gene models identified from RNA-Seq data, including incorrect predictions of introns, incorrect intron splice sites, novel introns and other incorrect annotations. B. One example of a gene with incorrect intron predictions is shown. Two introns were annotated in gene FGSG_01636 in the Broad F. graminearum database, but RNA-Seq data clearly showed that the second intron is absent. C. Three genes with incorrect intron predictions were selected for confirmation by RT-PCR. For each gene, one genomic DNA template and two cDNA templates (one is from wt PH-1, one is from mutant ebr1) were tested. Primers were designed flanking the intronic regions, as shown in B. Bands with identical size were amplified from both genomic DNA and cDNA template for all three genes, indicating that predicted introns do not exist. The correctly annotated gene FGSG_10264 was used as a control.