TY - CONF A1 - Gorbushina, Anna T1 - Some like it on the rocks: mineral-weathering fungi with manifold protective pigments N2 - The interface between materials and the environment is populated by microorganisms which organize on surfaces to form specially adapted and resistant microbial associations, so-called biofilms. Surface-bound biofilm growth is associated with substantial secretion of metabolic products that can significantly influence material corrosion. Here, a group of material-inhabiting fungi are presented, which colonize and weather air-exposed materials such as building façades, roofs and solar systems. A selected model fungus is used to carry out genetic and molecular biology experiments, which are microbiologically cutting-edge and allow for the development of novel time-saving techniques for material testing. The degree of material damage under the influence of this model fungus shows a measure of the aggressiveness of the biofilm under defined environmental conditions - and thus explores fundamental components of the interaction between material and biofilm. T2 - GZMB-Kolloquium Mikrobiologie/Genetik CY - Göttingen, Germany DA - 05.07.2016 KW - Microcolonial fungi KW - Sub-aerial biofilms KW - Knufia petricola PY - 2016 AN - OPUS4-37381 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Erdmann, Eileen A1 - Nitsche, Sarah A1 - Gorbushina, Anna A1 - Schumacher, Julia T1 - Genetic Engineering of the Rock Inhabitant Knufia petricola Provides Insight Into the Biology of Extremotolerant Black Fungi N2 - Black microcolonial fungi (Ascomycetes from Arthonio-, Dothideo-, and Eurotiomycetes) are stress-tolerant and persistent dwellers of natural and anthropogenic extreme habitats. They exhibit slow yeast-like or meristematic growth, do not form specialized reproduction structures and accumulate the black pigment 1,8-dihydroxynaphthalene (DHN) melanin in the multilayered cell walls. To understand how black fungi live, survive, colonize mineral substrates, and interact with phototrophs genetic methods are needed to test these functions and interactions. We chose the rock inhabitant Knufia petricola of the Chaetothyriales as a model for developing methods for genetic manipulation. Here, we report on the expansion of the genetic toolkit by more efficient multiplex CRISPR/Cas9 using a plasmid-based system for expression of Cas9 and multiple sgRNAs and the implementation of the three resistance selection markers genR (geneticin/nptII), baR (glufosinate/bar), and suR (chlorimuron ethyl/sur). The targeted integration of expression constructs by replacement of essential genes for pigment synthesis allows for an additional color screening of the transformants. The black-pink screening due to the elimination of pks1 (melanin) was applied for promoter studies using GFP fluorescence as reporter. The black-white screening due to the concurrent elimination of pks1 and phs1 (carotenoids) allows to identify transformants that contain the two expression constructs for co-localization or bimolecular fluorescence complementation (BiFC) studies. The co-localization and interaction of the two K. petricola White Collar orthologs were demonstrated. Two intergenic regions (igr1, igr2) were identified in which expression constructs can be inserted without causing obvious phenotypes. Plasmids of the pNXR-XXX series and new compatible entry plasmids were used for fast and easy generation of expression constructs and are suitable for a broad implementation in other fungi. This variety of genetic tools is opening a completely new perspective for mechanistic and very detailed study of expression, functioning and regulation of the genes/proteins encoded by the genomes of black fungi. KW - Microcolonial fungi KW - DHN melanin KW - Cloning vectors KW - Genetics PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-546192 DO - https://doi.org/10.3389/ffunb.2022.862429 SN - 2673-6128 VL - 3 PB - Frontiers Media CY - Lausanne AN - OPUS4-54619 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - C., Ruibal A1 - L., Selbmann A1 - Serap, Avci A1 - Martin-Sanchez, Pedro Maria A1 - Gorbushina, Anna ED - Gorbushina, Anna T1 - Roof-Inhabiting Cousins of Rock-Inhabiting Fungi: Novel Melanized Microcolonial Fungal Species from Photocatalytically Reactive Subaerial Surfaces N2 - Subaerial biofilms (SAB) are an important factor in weathering, biofouling, and biodeterioration of bare rocks, building materials, and solar panel surfaces. The realm of SAB is continually widened by modern materials, and the settlers on these exposed solid surfaces always include melanized, stress-tolerant microcolonial ascomycetes. After their first discovery on desert rock surfaces, these melanized chaetothyrialean and dothidealean ascomycetes have been found on Mediterranean monuments after biocidal treatments, Antarctic rocks and solar panels. New man-made modifications of surfaces (e.g., treatment with biocides or photocatalytically active layers) accommodate the exceptional stress-tolerance of microcolonial fungi and thus further select for this well-protected ecological group. Melanized fungal strains were isolated from a microbial community that developed on highly photocatalytic roof tiles after a long-term environmental exposure in a maritime-influenced region in northwestern Germany. Four of the isolated strains are described here as a novel species, Constantinomyces oldenburgensis, based on multilocus ITS, LSU, RPB2 gene phylogeny. Their closest relative is a still-unnamed rock-inhabiting strain TRN431, here described as C. patonensis. Both species cluster in Capnodiales, among typical melanized microcolonial rock fungi from different stress habitats, including Antarctica. These novel strains flourish in hostile conditions of highly oxidizing material surfaces, and shall be used in reference procedures in material testing. KW - Microcolonial fungi KW - Multilocus phylogeny KW - Photocatalytic surfaces KW - Subaerial biofilms KW - Stress tolerance KW - Constantinomyces PY - 2018 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-455182 DO - https://doi.org/10.3390/life8030030 VL - 8 IS - 3 SP - 30 EP - 44 PB - MDPI CY - Basel, Schweiz AN - OPUS4-45518 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Noack-Schönmann, Steffi A1 - Spagin, Olga A1 - Gründer, Klaus-Peter A1 - Breithaupt, Mathias A1 - Günther, Achim A1 - Muschik, Bernd A1 - Gorbushina, Anna T1 - Sub-aerial biofilms as blockers of solar radiation: spectral properties as tools to charcterise material-relevant microbial growth N2 - Sub-aerial biofilms (SABs) are ubiquitous microbial communities that develop at the interface between hard surfaces and the atmosphere. Inherent SAB 'core-settlers' include phototrophic algae, cyanobacteria, heterotrophic bacteria and microcolonial fungi (MCF). SABs do not simply cover hard surfaces; they interact with them in myriads of ways and bind to the underlying substrate. Secretion of extracellular mucilage aids adhesion, while organic acids and acidic polysaccharides weather the surface. As protection against solar radiation, many members of the SAB consortia produce shielding pigments while the phototrophic inhabitants are laden with photosynthetic pigments. All absorb light of many wavelengths and in addition, the cells themselves scatter light. Both effects change the spectra of incoming radiation (including wavelengths that are converted to electricity by photovoltaic cells) and decrease its intensity. To quantify these effects on SABs as complex entities of organisms and pigments, we measured the spectral properties of model and natural biofilms transferred to glass. Here we show that SABs growing on solar panels and other substrates scatter incident radiation between 250 nm up to 1800 nm and block up to 70% of its transmission. Model biofilms have the advantage that their microbial components can be 'tuned' to resemble natural ones of different compositions thus providing a novel materials-testing tool. KW - Material-colonising microorganisms KW - Microcolonial fungi KW - Glass biodeterioration KW - Solar panels KW - Biofilm-impaired transmittance of radiation PY - 2014 DO - https://doi.org/10.1016/j.ibiod.2013.09.020 SN - 0964-8305 VL - 86 SP - 286 EP - 293 PB - Elsevier CY - Barking AN - OPUS4-30128 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - 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 - CONF A1 - Gorbushina, Anna T1 - Some like it on the rocks: mineral-weathering fungi with manifold protective pigments N2 - The interface between materials and the environment is populated by microorganisms which organize on surfaces to form specially adapted and resistant microbial associations, so-called biofilms. Surface-bound biofilm growth is associated with substantial secretion of metabolic products that can significantly influence material corrosion. Here, a group of material-inhabiting fungi are presented, which colonize and weather air-exposed materials such as building façades, roofs and solar systems. A selected model fungus is used to carry out genetic and molecular biology experiments, which are microbiologically cutting-edge and allow for the development of novel time-saving techniques for material testing. The degree of material damage under the influence of this model fungus shows a measure of the aggressiveness of the biofilm under defined environmental conditions - and thus explores fundamental components of the interaction between material and biofilm. T2 - Kolloquium Bodenkunde CY - Göttingen, Germany DA - 06.07.2016 KW - Microcolonial fungi KW - Sub-aerial biofilms KW - Knufia petricola PY - 2016 AN - OPUS4-37382 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CHAP A1 - Knabe, Nicole A1 - Gorbushina, Anna ED - Gurtler, V. ED - Trevors, J. T. T1 - Territories of rock-inhabiting fungi: Survival on and alteration of solid air-exposed surfaces N2 - Subaerial biofilms that are omnipresent at the interface between all solid substrates and the atmosphere are composed of a unique and widespread group of ascomycetes called rock-inhabiting fungi or microcolonial fungi (MCF), typically in communities with other microorganisms. While subaerial biofilms in toto have important roles in mineral weathering and biodeterioration of materials, methodological approaches to subaerial biofilm communities are diverse and frequently focussed on MCF. Here, we review the historical development of the research methods applied in the field and consider perspectives to increase our understanding of the biofilm-induced changes of solid substrate surfaces. KW - Biologically induced mineral weathering KW - Geobiology KW - Microcolonial fungi KW - Subaerial biofilm KW - Symbiosis PY - 2018 UR - https://linkinghub.elsevier.com/retrieve/pii/S0580951718300047 SN - 9780128146040 DO - https://doi.org/10.1016/bs.mim.2018.06.001 VL - 45 SP - Chapter 6, 145 EP - 169 PB - Elsevier AN - OPUS4-47181 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Gorbushina, Anna A1 - Broughton, W.J. T1 - Microbiology of the atmosphere-rock interface: How biological interactions and physical stresses modulate a sophisticated microbial ecosystem N2 - Life at the atmosphere-lithosphere boundary is an ancient terrestrial niche that is sparsely covered by thin subaerial biofilms. The microbial inhabitants of these biofilms (a) have adapted to all types of terrestrial/subaerial stresses (e.g., desiccation, extreme temperatures, low nutrient availability, intense solar radiation), (b) interact with minerals that serve as both a dwelling and a source of mineral nutrients, and (c) provoke weathering of rocks and soil formation. Subaerial communities comprise heterotrophic and phototrophic microorganisms that support each other's lifestyle. Major lineages of eubacteria associated with the early colonization of land (e.g., Actinobacteria, Cyanobacteria) are present in these habitats along with eukaryotes such as microscopic green algae and ascomycetous fungi. The subaerial biofilm inhabitants have adapted to desiccation, solar radiation, and other environmental challenges by developing protective, melanized cell walls, assuming microcolonial architectures and symbiotic lifestyles. How these changes occurred, their significance in soil formation, and their potential as markers of climate change are discussed below. KW - Cyanobacteria KW - Geobiology KW - Microcolonial fungi KW - Rock inhabiting KW - Fungi-phototroph symbioses KW - Soil formation KW - Subaerial biofilms KW - Weathering PY - 2009 DO - https://doi.org/10.1146/annurev.micro.091208.073349 SN - 0066-4227 VL - 63 SP - 431 EP - 450 PB - Annual Reviews Inc. CY - Palo Alto, CA, USA AN - OPUS4-20243 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 - TY - CHAP A1 - Krumbein, W. A1 - Gorbushina, Anna ED - Watt, J. ED - Tidblad, J. ED - Kucera, V. ED - Hamilton, R. T1 - Some aspects of biological weathering and air pollution KW - Cyanobacteria KW - Geobiology KW - Microcolonial fungi KW - Rock inhabiting fungi-phototroph symbioses KW - Soil formation KW - Subaerial biofilms KW - Weathering PY - 2009 SN - 978-0-387-84892-1 DO - https://doi.org/10.1007/978-0-387-84893-8_5 IS - Chapter 5 SP - 127 EP - 145 PB - Springer Science + Business Media AN - OPUS4-20886 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -