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Deletion of gene clusters to identify novel secondary metabolites in Fusarium graminearum

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  • CGRB Fall Conference 2015
  • Previously we generated Fusarium graminearum mutants in which the gene for a histone methyltransferase (kmt6) was deleted [4]. In normal cells, this histone mark, trimethylated histone H3 lysine 27 (H3K27me3), is responsible for gene silencing or downregulation of ~25% of all genes. Many of these genes are in regions that are enriched for secondary metabolite gene clusters, compounds not required for primary growth but often associated with survival in hostile environments. Many secondary metabolites have been used as useful bioactive compounds or as antibiotics [2]. This is a promising approach as bacteria develop resistance to new compounds more quickly [1] In the kmt6 mutant many compounds were accumulating compared to wild type. As nearly 2,000 genes are upregulated or newly expressed, it is unclear which specific genes are associated with previously known or novel products. Our strategy is to delete key genes required for the production of metabolites in the kmt6 mutant to assign gene function to almost 650 overexpressed genes in 67 gene clusters. As proof‐of‐principle experiment, we focused on five clusters, replacing each cluster with a selectable maker that conveys antibiotic resistance. We validated transformants by PCR and Southern blots to show successful deletion of four of the five loci. The next step will be to analyze the metabolite profile of these mutants.
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  • This work was funded by grants from the NSF and NIH to the Freitag lab, with assistance from the CGRB.
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