TY - JOUR A1 - Nai, Corrado A1 - Wong, Helen A1 - Pannenbecker, A. A1 - Broughton, William J. A1 - Benoit, I. A1 - De Vries, R.P. A1 - Gueidan, C. A1 - Gorbushina, Anna T1 - Nutritional physiology of a rock-inhabiting, model microcolonial fungus from an ancestral lineage of the chaetothyriales (ascomycetes) N2 - Rock-inhabiting black fungi [also microcolonial or meristematic fungi (MCF)] are a phylogenetically diverse group of melanised ascomycetes with distinctive morphological features that confer extensive stress tolerance and permit survival in hostile environments. The MCF strain A95 Knufia petricola (syn. Sarcinomyces petricola) belongs to an ancestral lineage of the order Chaetothyriales (class Eurotiomycetes). K. petricola strain A95 is a rock-inhabiting MCF and its growth requirements were studied using the 96-well plate-based Biolog™ System under ~1070 different conditions (osmotic stress, pH growth optima, growth factor requirements and nutrient catabolism). A95 is an osmotolerant, oligotrophic MCF that grows best around pH 5. Remarkably, A95 shows metabolic activity in the absence of added nitrogen, phosphorus or sulphur. Correlations could be drawn between the known nutrient requirements of A95 and what probably is available in sub-aerial systems (rock and other material surfaces). Detailed knowledge of A95's metabolic requirements allowed formulation of a synthetic medium that supports strong fungal growth. KW - Knufia petricola (syn. Sarcinomyces petricola) KW - A95 KW - Chaetothyriales (Eurotiomycetes) KW - Microcolonial fungi KW - Melanised rock-inhabiting fungi KW - Biolog™ system KW - Physiological characterisation PY - 2013 DO - https://doi.org/10.1016/j.fgb.2013.04.001 SN - 1087-1845 SN - 0147-5975 SN - 1096-0937 VL - 56 SP - 54 EP - 66 PB - Elsevier CY - Amsterdam [u.a.] AN - OPUS4-28865 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Erdmann, Eileen A. A1 - Brandhorst, Antonia K. M. A1 - Gorbushina, Anna A1 - Schumacher, Julia T1 - The Tet‑on system for controllable gene expression in the rock‑inhabiting black fungus Knufia petricola N2 - Knufia petricola is a black fungus that colonizes sun-exposed surfaces as extreme and oligotrophic environments. As ecologically important heterotrophs and biofilm-formers on human-made surfaces, black fungi form one of the most resistant groups of biodeteriorating organisms. Due to its moderate growth rate in axenic culture and available protocols for its transformation and CRISPR/Cas9-mediated genome editing, K. petricola is used for studying the morpho-physiological adaptations shared by extremophilic and extremotolerant black fungi. In this study, the bacteria-derived tetracycline (TET)-dependent promoter (Tet-on) system was implemented to enable controllable gene expression in K. petricola. The functionality i.e., the dose-dependent inducibility of TET-regulated constructs was investigated by using GFP fluorescence, pigment synthesis(melanin and carotenoids) and restored uracil prototrophy as reporters. The newly generated cloning vectors containing the Tet-on construct, and the validated sites in the K. petricola genome for color-selectable or neutral insertion of expression constructs complete the reverse genetics toolbox. One or multiple genes can be expressed on demand from different genomic loci or from a single construct by using 2A self-cleaving peptides, e.g., for localizing proteins and protein complexes in the K. petricola cell or for using K. petricola as host for the expression of heterologous genes. KW - Microcolonial fungi KW - Inducible promoter KW - Bimolecular fluorescence complementation KW - 2A peptide KW - CRISPR/ Cas9-mediated genome editing PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-607672 DO - https://doi.org/10.1007/s00792-024-01354-2 VL - 28 IS - 38 SP - 1 EP - 13 PB - Springer Nature AN - OPUS4-60767 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -