Bioinformatics Prediction of Polyketide Synthase Gene Clusters from Mycosphaerella fijiensis

PLoS One. 2016 Jul 7;11(7):e0158471. doi: 10.1371/journal.pone.0158471. eCollection 2016.

Abstract

Mycosphaerella fijiensis, causal agent of black Sigatoka disease of banana, is a Dothideomycete fungus closely related to fungi that produce polyketides important for plant pathogenicity. We utilized the M. fijiensis genome sequence to predict PKS genes and their gene clusters and make bioinformatics predictions about the types of compounds produced by these clusters. Eight PKS gene clusters were identified in the M. fijiensis genome, placing M. fijiensis into the 23rd percentile for the number of PKS genes compared to other Dothideomycetes. Analysis of the PKS domains identified three of the PKS enzymes as non-reducing and two as highly reducing. Gene clusters contained types of genes frequently found in PKS clusters including genes encoding transporters, oxidoreductases, methyltransferases, and non-ribosomal peptide synthases. Phylogenetic analysis identified a putative PKS cluster encoding melanin biosynthesis. None of the other clusters were closely aligned with genes encoding known polyketides, however three of the PKS genes fell into clades with clusters encoding alternapyrone, fumonisin, and solanapyrone produced by Alternaria and Fusarium species. A search for homologs among available genomic sequences from 103 Dothideomycetes identified close homologs (>80% similarity) for six of the PKS sequences. One of the PKS sequences was not similar (< 60% similarity) to sequences in any of the 103 genomes, suggesting that it encodes a unique compound. Comparison of the M. fijiensis PKS sequences with those of two other banana pathogens, M. musicola and M. eumusae, showed that these two species have close homologs to five of the M. fijiensis PKS sequences, but three others were not found in either species. RT-PCR and RNA-Seq analysis showed that the melanin PKS cluster was down-regulated in infected banana as compared to growth in culture. Three other clusters, however were strongly upregulated during disease development in banana, suggesting that they may encode polyketides important in pathogenicity.

MeSH terms

  • Ascomycota / enzymology
  • Ascomycota / genetics*
  • Computational Biology*
  • Fungal Proteins / genetics*
  • Fusarium / genetics
  • Genome, Fungal
  • Likelihood Functions
  • Multigene Family*
  • Musa / microbiology
  • Open Reading Frames
  • Peptide Synthases / genetics
  • Phylogeny
  • Plant Diseases / microbiology
  • Polyketide Synthases / genetics*
  • Polyketides
  • Protein Domains
  • Sequence Analysis, RNA

Substances

  • Fungal Proteins
  • Polyketides
  • Polyketide Synthases
  • Peptide Synthases
  • non-ribosomal peptide synthase

Grants and funding

This work was funded by a gift from Dole Food Company to MED and by graduate fellowships from the National Science Foundation and National Institutes of Health to RDN. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.