Protoplast-mediated transformation of Madurella mycetomatis using hygromycin resistance as a selection marker

PLoS Negl Trop Dis. 2024 Apr 5;18(4):e0012092. doi: 10.1371/journal.pntd.0012092. eCollection 2024 Apr.

Abstract

Madurella mycetomatis is the main cause of mycetoma, a chronic granulomatous infection for which currently no adequate therapy is available. To improve therapy, more knowledge on a molecular level is required to understand how M. mycetomatis is able to cause this disease. However, the genetic toolbox for M. mycetomatis is limited. To date, no method is available to genetically modify M. mycetomatis. In this paper, a protoplast-mediated transformation protocol was successfully developed for this fungal species, using hygromycin as a selection marker. Furthermore, using this method, a cytoplasmic-GFP-expressing M. mycetomatis strain was created. The reported methodology will be invaluable to explore the pathogenicity of M. mycetomatis and to develop reporter strains which can be useful in drug discovery as well as in genetic studies.

MeSH terms

  • Cinnamates / pharmacology
  • Drug Resistance, Fungal / genetics
  • Hygromycin B* / analogs & derivatives
  • Hygromycin B* / pharmacology
  • Madurella* / drug effects
  • Madurella* / genetics
  • Mycetoma / drug therapy
  • Mycetoma / microbiology
  • Protoplasts*
  • Transformation, Genetic*

Substances

  • hygromycin A

Grants and funding

This work was supported by the H2020 Marie Sklodowska Curie-COFUND program LEaDing Fellows, grant number 707404 awarded to SdP and Aspasia grant 015.013.033 from the Dutch Research Council and an EUR fellowship of the Erasmus University both awarded to WvdS The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.