A molecular toolbox for manipulating Eremothecium coryli

Microbiol Res. 2007;162(4):299-307. doi: 10.1016/j.micres.2007.05.008. Epub 2007 Aug 22.

Abstract

The genus Eremothecium contains dimorphic and filamentous fungal species, most notably Eremothecium sinecaudum (Holleya sinecauda), a dimorphic plant pathogen, which was isolated from mustard seeds, and Eremothecium gossypii (Ashbya gossypii), a filamentous fungus, which is well known for its ability to produce riboflavin. In this study, we present the initial molecular characterization of another Eremothecium species classified as Eremothecium coryli. E.coryli is a dimorphic fungus. We have developed, based on previously described reagents, a transformation system for E. coryli using kanMX and NATMX3 as dominant selectable marker genes on freely replicating plasmids conferring resistance to the antibiotics G418 and nourseothricin, respectively. As reporter genes we could introduce lacZ and GFP, which were controlled either by the AgTEF1 promoter or by regulatable MET promoters derived from the A. gossypii and Saccharomyces cerevisiae MET3 genes. These newly established tools will allow a detailed comparison of different growth modes in filamentous or dimorphic species within the genus Eremothecium.

MeSH terms

  • Antifungal Agents / pharmacology
  • Artificial Gene Fusion
  • Drug Resistance, Fungal / genetics
  • Genes, Reporter
  • Genetic Vectors
  • Gentamicins / pharmacology
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics
  • Microbiological Techniques / methods*
  • Molecular Biology / methods*
  • Plasmids
  • Promoter Regions, Genetic
  • Saccharomyces cerevisiae / genetics
  • Saccharomycetales / cytology
  • Saccharomycetales / drug effects
  • Saccharomycetales / genetics*
  • Saccharomycetales / physiology
  • Streptothricins / pharmacology
  • Transformation, Genetic
  • beta-Galactosidase / biosynthesis
  • beta-Galactosidase / genetics

Substances

  • Antifungal Agents
  • Gentamicins
  • Green Fluorescent Proteins
  • Streptothricins
  • antibiotic G 418
  • beta-Galactosidase