Zitieren Sie bitte immer diesen URN: urn:nbn:de:kobv:b43-518601
An advanced genetic toolkit for exploring the biology of the rock‑inhabiting black fungus Knufia petricola
- Microcolonial black fungi are a group of ascomycetes that exhibit high stress tolerance, yeast-like growth and constitutive melanin formation. They dominate a range of hostile natural and man-made environments, from desert rocks and salterns to dishwashers, roofs and solar panels. Due to their slow growth and a lack of genetic tools, the underlying mechanisms of black fungi’s phenotypic traits have remained largely unexplored. We chose to address this gap by genetically engineering the rock-inhabiting fungus Knufia petricola (Eurotiomycetes, Chaetothyriales), a species that exhibits all characteristics of black fungi. A cell biological approach was taken by generating K. petricola strains expressing green or red fluorescent protein variants. By applying: (1) traditional gene replacement; (2) gene editing and replacement via plasmid-based or ribonucleoprotein (RNP)-based CRISPR/Cas9, and (3) silencing by RNA interference (RNAi), we constructed mutants in the pathways leading to melanin,Microcolonial black fungi are a group of ascomycetes that exhibit high stress tolerance, yeast-like growth and constitutive melanin formation. They dominate a range of hostile natural and man-made environments, from desert rocks and salterns to dishwashers, roofs and solar panels. Due to their slow growth and a lack of genetic tools, the underlying mechanisms of black fungi’s phenotypic traits have remained largely unexplored. We chose to address this gap by genetically engineering the rock-inhabiting fungus Knufia petricola (Eurotiomycetes, Chaetothyriales), a species that exhibits all characteristics of black fungi. A cell biological approach was taken by generating K. petricola strains expressing green or red fluorescent protein variants. By applying: (1) traditional gene replacement; (2) gene editing and replacement via plasmid-based or ribonucleoprotein (RNP)-based CRISPR/Cas9, and (3) silencing by RNA interference (RNAi), we constructed mutants in the pathways leading to melanin, carotenoids, uracil and adenine. Stable single and double mutants were generated with homologous recombination (HR) rates up to 100%. Efficient, partially cloning-free strategies to mutate multiple genes with or without resistance cassettes were developed. This state-of-the-art genetic toolkit, together with the annotated genome sequence of strain A95, firmly established K. petricola as a model for exploring microcolonial black fungi.…
Autor*innen: | Oliver Voigt, Nicole Knabe, Sarah Nitsche, Eileen Erdmann, Julia SchumacherORCiD, Anna GorbushinaORCiD |
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Dokumenttyp: | Zeitschriftenartikel |
Veröffentlichungsform: | Verlagsliteratur |
Sprache: | Englisch |
Titel des übergeordneten Werkes (Englisch): | Scientific Reports |
Jahr der Erstveröffentlichung: | 2020 |
Organisationseinheit der BAM: | 4 Material und Umwelt |
4 Material und Umwelt / 4.0 Abteilungsleitung und andere | |
Veröffentlichende Institution: | Bundesanstalt für Materialforschung und -prüfung (BAM) |
Verlag: | Springer Nature |
Jahrgang/Band: | 10 |
Ausgabe/Heft: | 1 |
Erste Seite: | 22021 |
DDC-Klassifikation: | Technik, Medizin, angewandte Wissenschaften / Ingenieurwissenschaften / Sanitär- und Kommunaltechnik; Umwelttechnik |
Freie Schlagwörter: | Biodeterioration; CRISPR/Cas9; Fluorescent microscopy; RNA interference; Subaerial biofilms |
Themenfelder/Aktivitätsfelder der BAM: | Umwelt |
Umwelt / Umwelt-Material-Interaktionen | |
DOI: | 10.1038/s41598-020-79120-5 |
URN: | urn:nbn:de:kobv:b43-518601 |
Verfügbarkeit des Dokuments: | Datei für die Öffentlichkeit verfügbar ("Open Access") |
Lizenz (Deutsch): | Creative Commons - CC BY - Namensnennung 4.0 International |
Datum der Freischaltung: | 17.12.2020 |
Referierte Publikation: | Ja |
Datum der Eintragung als referierte Publikation: | 04.01.2021 |
Schriftenreihen ohne Nummerierung: | Wissenschaftliche Artikel der BAM |