New capabilities for Mycosphaerella graminicola research

Mol Plant Pathol. 2010 Sep;11(5):691-704. doi: 10.1111/j.1364-3703.2010.00629.x.

Abstract

Mycosphaerella graminicola is a major pathogen of wheat worldwide, causing Septoria leaf blotch disease. Targeted gene disruption in M. graminicola, by Agrobacterium tumefaciens-mediated transformation, has become an established functional genomics tool for M. graminicola research in recent years. However, in order to advance research into this economically important pathogen, further functional genomics tools need to be developed. Here, we report three new capabilities for M. graminicola research: (i) two selectable markers have been shown to work robustly in M. graminicola, namely G418 and the fungicide carboxin; (ii) the generation of a strain of M. graminicola in which the KU70 (MUS-51) homologue has been disrupted; in this strain, homologous recombination efficiencies increased to more than 95%, whilst maintaining wild-type growth in vitro and full pathogenicity on wheat leaves; (iii) the ability to efficiently target and generate precise mutations of specific genes in the genomic context in M. graminicola. In addition, the insertion of the E198A mutation into the beta-tubulin gene (MgTUB1), conferring resistance to the fungicide benomyl, suggests that this mutant allele may provide an additional selectable marker. The collective use of these tools will permit further advancements in our knowledge of the biology and pathogenicity of this important plant pathogen.

MeSH terms

  • Alleles
  • Analysis of Variance
  • Antigens, Nuclear / chemistry
  • Antigens, Nuclear / genetics
  • Ascomycota / drug effects
  • Ascomycota / genetics*
  • Ascomycota / growth & development
  • Benomyl / pharmacology
  • Blotting, Southern
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • Drug Resistance, Fungal / drug effects
  • Drug Resistance, Fungal / genetics
  • Gene Deletion
  • Gene Targeting
  • Genetic Loci / genetics
  • Genetic Markers
  • Genetic Research*
  • Ku Autoantigen
  • Mutagenesis, Insertional / genetics
  • Phenotype
  • Point Mutation / genetics
  • Recombination, Genetic / drug effects
  • Recombination, Genetic / genetics
  • Reproducibility of Results
  • Selection, Genetic / drug effects
  • Sequence Homology, Amino Acid
  • Transformation, Genetic / drug effects
  • Tubulin / genetics

Substances

  • Antigens, Nuclear
  • DNA-Binding Proteins
  • Genetic Markers
  • Tubulin
  • Ku Autoantigen
  • Benomyl