TY - JOUR A1 - Mallikarjun, Jhenkhar A1 - Gorbushina, Anna A1 - Kuzyakov, Yakov A1 - Koester, Moritz A1 - Castro, Rodrigo A1 - Yudina, Anna A1 - Abdallah, Khaled A1 - Matus, Francisco J. A1 - Dippold, Michaela A. T1 - Post-fire recovery of temperate and mediterranean ecosystems: An interplay between fire severity, soil nutrients, and vegetation from early-stage to decadal-scale dynamics N2 - Wildfires strongly alter soil properties, which in turn affect ecosystem recovery over extended periods, though long-term impacts are less certain. This study investigated a 14-year post-fire chronosequence in Chile’s mediterranean and temperate humid forests, revealing ecosystem-specific soil properties and nutrient recovery mechanisms. By analysing sites at successional stages, the chronosequence approach assessed temporal changes and ecosystem recovery, revealing long-term wildfire effects on soil dynamics and nutrients recovery. Wildfires raised soil bulk density to 0.9 g cm−3 in humid temperate and 1.2 g cm−3 in mediterranean ecosystems. Mediterranean soils experienced greater compaction from organic matter loss, soil aggregate destruction, ash-clogged pores, and topsoil erosion. Soil texture shifts were ecosystem-dependent: mediterranean soils increased 10–12 % in clay and silt through ash redistribution and aggregation, while temperate soils saw sand content rise by 0.74 % and 0.32 % yearly at 0–5 and 5–10 cm depths from thermal disaggregation and erosion. Ground vegetation recovers quickly, but physical soil properties like bulk density require over 14 years to return to pre-fire conditions. In humid temperate forests, ash input initially increased soil pH (4.8 to 5.8), reducing acidity, mitigating aluminium toxicity, while increasing nutrient availability. Base cation stocks increased in mediterranean woodlands (e.g., Ca: up to 0.41 Mg ha−1 y−1) due to ash retention, lower leaching, and ash infiltration into subsoil. Nutrient stocks in humid forests recovered slowly (Ca: 0.087–0.13 Mg ha−1 y−1) due to rainfall-driven leaching and low subsoil reserves. Carbon and N losses were restricted to the litter horizon in temperate forests, recovering via fire-resistant tree inputs, whereas mediterranean soils suffered severe C and N depletion from vegetation loss, erosion, and low N fixation. Fire effects and recovery are ecosystem-specific, shaped by landscape, geology, hydrology, and vegetation resilience. Understanding how fire regimes affect soil and nutrient recovery is vital for improving projections in fire-prone regions. KW - Historical fire & Wildfire KW - Nutrients Ecosystem recovery & Vegetation succession KW - Chronosequence approach KW - Losses of nutrients & Nutrients stocks KW - Wildfire PY - 2025 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-642918 DO - https://doi.org/10.1016/j.catena.2025.109431 SN - 0341-8162 VL - 260 SP - 1 EP - 13 PB - Elsevier B.V. AN - OPUS4-64291 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Wachsender Faktor der Energiewende: Biofilme auf Photovoltaikanlagen T2 - Vortrag bei der Universität Oldenburg CY - Oldenburg, Germany DA - 2014-04-29 PY - 2014 AN - OPUS4-31302 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Co-evolution of rock surfaces and their biofilms T2 - Einweihung der HELGES-Labore für die Geochemie der Erdoberfläche am Deutschen GeoForschungsZentrum CY - Potsdam, Germany DA - 2013-06-11 PY - 2013 AN - OPUS4-28884 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Martin-Sanchez, Pedro Maria A1 - Gorbushina, Anna A1 - Toepel, Jörg T1 - Quantification of microbial load in diesel storage tanks using culture- and qPCR-based approaches N2 - Microbial contamination of fuels, associated with a wide variety of bacteria and fungi, leads to decreased product quality and can compromise equipment performance by biofouling and microbiologically influenced corrosion of pipelines and storage tanks. Detection and quantification of biomass are critical in monitoring fuel systems for an early detection of microbial outbreaks. The aims of this study are (i) to quantify bacterial and fungal contamination in samples from diesel storage tanks of petrol stations, using both culture dependent- and culture independent (qPCR) approaches, and (ii) to analyse the diversity of cultivable diesel-contaminating microorganisms with the purpose to create a strain collection for further use in biodeterioration experiments. Both methodological approaches revealed a high microbial contamination in all studied samples, with the bacterial load being much higher than the fungal load. The diversity of cultivable microorganisms was rather low. Based on criteria of abundance and fuel degradation potential, the most relevant microorganisms were identified as bacteria of genera Bacillus, Citrobacter, Burkholderia and Acetobacter, the filamentous fungi Paecilomyces variotii and Pseudallescheria boydii, and a Dipodascaceae yeast. Furthermore the validity and utility of qPCR-based methods are discussed. KW - Fuel contamination KW - Biofouling KW - Cultivation KW - Real-time qPCR KW - Bacteria KW - Fungi PY - 2016 DO - https://doi.org/10.1016/j.ibiod.2016.04.009 SN - 0964-8305 SN - 1879-0208 VL - 126 SP - 216 EP - 223 PB - Elsevier Ltd. AN - OPUS4-41260 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Studying colonization of stone surfaces – what can we learn from model biofilms growing in flow-through chambers? N2 - Soil formation on weathering rock surfaces is intrinsically connected with the development of primary microbial colonization at the atmosphere-lithosphere interface. A great number and variety of microorganisms is involved in these microbial communities, which are dominated by fungi, algae, cyanobacteria and heterotrophic bacteria. Rock-inhabiting life is ubiquitous on rock surfaces all around the world, but the laws of its establishment, and more important, quantification of its biodeterioration and geological input are possible only in well-controlled and simplified laboratory models. Here we would like to compare two model rock biofilm consisting of the heterotrophic and the phototrophic interacting partners. In the present work the growth of these model biofilms on diverse materials with different physical and chemical properties was investigated under well-controlled laboratory conditions. To clarify the role of environmental factors, the parameters temperature, light intensity and relative humidity were varied in growth test series. For an accelerated substrate colonization and to increase the biomass yield different flow-through chambers systems with semi-continuous cultures have been applied, simulating weathering conditions like flooding, desiccation and nutrient input. The biofilm development was studied by (i) confocal laser scanning and electron microscopy and (ii) qualitatively and quantitatively with respect to cell forms and biomass. A correlation between the presence of the model biofilm and mineral surface alteration as well as geochemical tracers of weathering were followed on various rock substrates (with differing geochemistry, porosity etc) exposed in another flow-through chamber, filled with crushed rock material. Under mentioned environmental conditions different types of flow-through chambers have been used and will be compared. T2 - Technoheritage CY - Cadiz, Spain DA - 21.05.2017 KW - Biofilm KW - Biodeterioration PY - 2017 AN - OPUS4-41136 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Life on the Rocks or Subaerial Biofilms T2 - Mikrobiologisches Kolloquium, Institut für Biologie an der Freien Universität, Fachbereich Biologie, Chemie, Pharamzie CY - Berlin, Germany DA - 2010-01-29 PY - 2010 AN - OPUS4-20864 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - How biological interactions and physical stresses influence fungal; biogeochemical impact T2 - Goldschmidt-Tagung 2009 CY - Davos, Switzerland DA - 2009-06-23 PY - 2009 AN - OPUS4-19981 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Life on the rocks: microscopic landscapes under the influence of microbes and atmosphere T2 - Vortrag CY - Potsdam, Germany DA - 2010-03-10 PY - 2010 AN - OPUS4-21104 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Material und Umwelt: Mikroben als Materialbesiedler T2 - GfKORR-Arbeitskreis "Mikrobiell beeinflusste Korrosion" der DECHEMA-Fachgruppe "Mikrobielle Materialzerstörung und Materialschutz" CY - Frankfurt am Main, Germany DA - 2011-10-21 PY - 2011 AN - OPUS4-24675 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Toepfer, I. A1 - Favet, J. A1 - Schulte, A. A1 - Schmölling, M. A1 - Butte, W. A1 - Triplett, E.W. A1 - Broughton, W.J. A1 - Gorbushina, Anna T1 - Pathogens as potential hitchhikers on intercontinental dust N2 - Desert dust seeds distant lands and waters with minerals as well as micro-organisms raising the question of whether this ancient phenomenon also spreads pathogens across the globe. Severe dust storms require strong winds blowing over land-masses that are largely devoid of vegetation, effectively limiting the scope for winds to raise pathogens into the air. Nevertheless, changing patterns of land-use, often driven by belligerency, result in refugees spreading to areas that were previously deemed barely habitable. With the help of the International Committee of the Red Cross, a number of sand/dust samples were collected from the Republic of Chad, some near refugee camps, others further removed from human influence. In parallel studies, we documented the micro-organisms present in these samples and used a number of the isolates here to test the effect of environmental constraints on their ability to survive intercontinental flight. We also added traditional pathogens to the palette of microbes and tested the effects of UV irradiation, desiccation and temperature on survival of both bacteria and fungi. A clear trend was obvious—those microbes that are coloured or able to form conidia or spores (in other words, those that are native to deserts) were well able to resist the imposed stresses. On the other hand, most pathogens were more sensitive to stresses than the environmental isolates. Toxin production in two species of Aspergillus was also investigated. Short-term desiccation (simulating environmental conditions during intercontinental travel) of sand amended with fungal spores containing sterigmatocystin leads to increased mycotoxin contents, but significant mycotoxin production was only possible under growth-permissive conditions, e.g. at higher humidity. It thus seems likely that an ever-decreasing fraction of the initial pathogen load survives as the dust recedes from its desert source and that those organisms that land on other continents are highly enriched in desert dwellers. KW - Chad KW - Desert sand KW - Sterigmatocystin KW - Aspergillus sydowii KW - A. versicolor PY - 2012 DO - https://doi.org/10.1007/s10453-011-9230-2 SN - 0393-5965 SN - 1573-3025 VL - 28 IS - 2 SP - 221 EP - 231 PB - Springer Science + Business Media B.V. CY - Dordrecht [u.a.] AN - OPUS4-24814 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Life on the rocks and other hard surfaces N2 - After their discovery on rock surfaces in cold and hot deserts, a polyphyletic group of ascomycetous black fungi was found to dominate a range of hostile environments – natural and man-made, from salterns to dishwashers, roofs and solar panels. Together with bacteria and algae they may establish subaerial biofilms and cause weathering of the surfaces they grow on. Their impressive survival abilities as well as their constitutive protective pigmentation and cluster-like microcolony organisation are similar in environmental isolates as well as in heat-tolerant opportunistic pathogens of animals and humans. We selected the rock-inhabiting fungus Knufia petricola (class Eurotiomycetes, order Chaetothyriales) that exhibits all the characteristics of microcolonial black fungi such as yeast-like cell growth, absence of reproductive structures and constitutive dihydroxynaphthalene (DHN) melanogenesis (Nai et al. 2013, Fungal Genet Biol). We developed protocols to efficiently generate and transform protoplasts resulting in stable homokaryotic transformants by targeting genes involved in pigment synthesis. The differences between the wild-type fungus and its melanin-deficient mutants were tested in geomicrobiological experiments and measured are now discussed in detail, with special accent on the possible effects of the mutation on EPS and other exuded substances. T2 - Departmental colloquium of the Tuscia University CY - Viterbo, Italy DA - 14.02.2020 KW - Ascomycetes KW - Fungal biofilms KW - Material surface colonisation KW - Genetics KW - Genomics KW - Extremophilic fungi PY - 2020 AN - OPUS4-50643 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - DNA EXTRACTION METHODS, AS WELL AS GENOME ANALYSIS OF SEVERAL BLACK FUNGI :Genomic and molecular characterization of a model ascomycete ancestral to mutualistic and pathogen-rich fungal lineages (A95 Sarcinomyces petricola, Chaetothyriales) T2 - Fourth meeting of the ISHAM working groups on Black Yeasts and chromoblastomycosis: “Hidden Danger, Bright Promise” CY - Curitiba, Brazil DA - 2011-12-01 PY - 2011 AN - OPUS4-26469 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Montero-Calasanz, M.C. A1 - Göker, M. A1 - Pötter, G. A1 - Rohde, M. A1 - Spröer, C. A1 - Schumann, P. A1 - Gorbushina, Anna A1 - Klenk, H.-P. T1 - Geodermatophilus arenarius sp. nov., a xerophilic actinomycete isolated from Saharan desert sand in Chad N2 - A novel Gram-positive, aerobic, actinobacterial strain, CF5/4T, was isolated in 2007 during an environmental screening of arid desert soil in Ouré Cassoni, Chad. The isolate grew best in a temperature range of 28–40 °C and at pH 6.0-8.5, with 0-1 % (w/v) NaCl, forming brown-coloured and nearly circular colonies on GYM agar. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The DNA G + C content of the novel strain was 75.9 mol %. The peptidoglycan contained meso-diaminopimelic acid as diagnostic diaminoacid. The main phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, diphosphatidylglycerol and a small amount of phosphatidylglycerol; MK-9(H4) was identified as the dominant menaquinone and galactose as diagnostic sugar. The major cellular fatty acids were branched-chain saturated acids: iso-C15:0 and iso-C16:0. The 16S rRNA gene showed 96.2–98.3 % sequence identity with the three members of the genus Geodermatophilus: G. obscurus (96.2 %), G. ruber (96.5 %), and G. nigrescens (98.3 %). Based on the chemotaxonomic results, 16S rRNA gene sequence analysis and DNA–DNA hybridization with the type strain of G. nigrescens, the isolate is proposed to represent a novel species, Geodermatophilus arenarius (type strain CF5/4T = DSM 45418T = MTCC 11413T = CCUG 62763T). KW - Xerophiles KW - Actinomycetes KW - Geodermatophilaceae KW - Taxonomy KW - Sahara desert PY - 2012 DO - https://doi.org/10.1007/s00792-012-0486-4 SN - 1431-0651 SN - 1433-4909 VL - 16 IS - 6 SP - 903 EP - 909 PB - Springer CY - Tokyo AN - OPUS4-26933 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Montero-Calasanz, M.d.C. A1 - Göker, M. A1 - Pötter, G. A1 - Rohde, M. A1 - Spröer, C. A1 - Schumann, P. A1 - Gorbushina, Anna A1 - Klenk, H.-P. T1 - Geodermatophilus africanus sp. nov., a halotolerant actinomycete isolated from Saharan desert sand N2 - A novel Gram-strain positive, aerobic, actinobacterial strain, designated CF11/1T, was isolated from a sand sample obtained in the Sahara Desert, Chad. The black-pigmented isolate was aerobic and exhibited optimal growth from 25 to 35 °C at pH 6.0–8.0 and with 0–8 % (w/v) NaCl, indicating that it is a halotolerant mesophile. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The G+C content in the genome was 74.4 mol%. The peptidoglycan contained meso-diaminopimelic acid as diagnostic diaminoacid. The main phospholipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and a minor fraction of phosphatidylglycerol; MK-9(H4) was the dominant menaquinone, and galactose was detected as a diagnostic sugar. The major cellular fatty acid was branched-chain saturated acid iso-C16:0. Analysis of 16S rRNA gene sequences showed 95.3–98.6 % pairwise sequence identity with the members of the genus Geodermatophilus. Based on phenotypic and chemotaxonomic properties, as well as phylogenetic distinctiveness, the isolate represents a novel species, Geodermatophilus africanus, with the type strain CF11/1T (DSM 45422 = CCUG 62969 = MTCC 11556). KW - Actinomycetes KW - Geodermatophilaceae KW - Taxonomy KW - Osmotolerant KW - Phenotype microarray PY - 2013 DO - https://doi.org/10.1007/s10482-013-9939-8 SN - 0003-6072 SN - 1572-9699 VL - 104 IS - 2 SP - 207 EP - 216 PB - Springer CY - Dordrecht AN - OPUS4-28868 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Bacterial Attachment to Poly[acrylonitrile-co-(2-methyl-2-propene-1-sulfonic acid)] T2 - MRS Spring Meeting & Exhibit 2013, Symposium NN: Multifunctional Biomaterials CY - San Francisco, CA, USA DA - 2013-04-01 PY - 2013 AN - OPUS4-28899 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Microbiota as the weathering engine T2 - Earth Shape: Earth Surface Shaping by Biota; Kick-off Meeting CY - GFZ, Potsdam, Germany DA - 2014-10-30 PY - 2014 AN - OPUS4-31921 LA - deu 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 - 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 - Schumacher, Julia A1 - Gorbushina, Anna T1 - Schwarze Pilze – Wüstenbesiedler finden neue Lebensräume N2 - Schwarze mikrokoloniale Pilze besiedeln zunehmend von Menschen geschaffene Habitate, wie schadstoffbelastete Böden, Statuen, Gebäudefassaden und Dächer. Sie verfärben und zersetzen die Oberflächen von anfälligen Materialien oder reduzieren die Lichtausbeute von Solaranlagen. Die Biologie dieser Pilze und ihre Relevanz für die Materialforschung stehen im Fokus unserer Studien an der Bundesanstalt für Materialforschung und -prüfung (BAM). Mit einer Kollektion schwarzer Pilze, die von Solaranlagen isoliert wurden, bringen wir klima- und materialrelevante Biodiversität in den Stammbaum des Lebens. Die Überlebensstrategien dieser Organismen versuchen wir mit molekularbiologischen und genetischen Untersuchungsansätzen zu entschlüsseln. KW - Pilze KW - Genetik KW - Diversität PY - 2021 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-541037 DO - https://doi.org/10.1007/s12268-021-1646-9 VL - 27 IS - 6 SP - 665 EP - 666 PB - Springer AN - OPUS4-54103 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Catanzaro, Ilaria A1 - Gerrits, Ruben A1 - Feldmann, Ines A1 - Gorbushina, Anna A1 - Onofri, Silvano A1 - Schumacher, Julia T1 - Deletion of the polyketide synthase‐encoding gene pks1 prevents melanization in the extremophilic fungus Cryomyces antarcticus N2 - Cryomyces antarcticus, a melanized cryptoendolithic fungus endemic to Antarctica, can tolerate environmental conditions as severe as those in space. Particularly, its ability to withstand ionizing radiation has been attributed to the presence of thick and highly melanized cell walls, which—according to a previous investigation—may contain both 1,8‐dihydroxynaphthalene (DHN) and L‐3,4 dihydroxyphenylalanine (L‐DOPA) melanin. The genes putatively involved in the synthesis of DHN melanin were identified in the genome of C. antarcticus. Most important is capks1 encoding a non‐reducing polyketide synthase (PKS) and being the ortholog of the functionally characterized kppks1 from the rock‐inhabiting fungus Knufia petricola. The co‐expression of CaPKS1 or KpPKS1 with a 4′‐phosphopantetheinyl transferase in Saccharomyces cerevisiae resulted in the formation of a yellowish pigment, suggesting that CaPKS1 is the enzyme providing the precursor for DHN melanin. To dissect the composition and function of the melanin layer in the outer cell wall of C. antarcticus, non‐melanized mutants were generated by CRISPR/Cas9‐mediated genome editing. Notwithstanding its slow growth (up to months), three independent non‐melanized Δcapks1 mutants were obtained. The mutants exhibited growth similar to the wild type and a light pinkish pigmentation, which is presumably due to carotenoids. Interestingly, visible light had an adverse effect on growth of both melanized wild‐type and non‐melanized Δcapks1 strains. Further evidence that light can pass the melanized cell walls derives from a mutant expressing a H2B‐GFP fusion protein, which can be detected by fluorescence microscopy. In conclusion, the study reports on the first genetic manipulation of C. antarcticus, resulting in non‐melanized mutants and demonstrating that the melanin is rather of the DHN type. These mutants will allow to elucidate the relevance of melanization for surviving extreme conditions found in the natural habitat as well as in space. KW - Astrobiology KW - Black fungi KW - CRISPR/Cas9 KW - DHN melanin KW - Cryptoendolithism PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-606599 DO - https://doi.org/10.1002/iub.2895 SN - 1521-6551 VL - 76 IS - 12 SP - 1072 EP - 1090 PB - Wiley AN - OPUS4-60659 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 - Abdallah, Khaled A1 - Stock, S. C. A1 - Heeger, Felix A1 - Koester, M. A1 - Nájera, F. A1 - Merino, C. A1 - Spielvogel, S. A1 - Gorbushina, Anna A1 - Kuzyakov, Y. A1 - Dippold, M. A. T1 - Nitrogen Gain and Loss Along an Ecosystem Sequence: From Semi-desert to Rainforest N2 - Plants and microorganisms, besides the climate, drive nitrogen (N) cycling in ecosystems. Our objective was to investigate N losses and N acquisition strategies along a unique ecosystem-sequence (ecosequence) ranging from arid shrubland through Mediterranean woodland to temperate rainforest. These ecosystems differ in mean annual precipitation, mean annual temperate, and vegetation cover, but developed on similar granitoid soil parent material, were addressed using a combination of molecular biology and soil biogeochemical tools. Soil N and carbon (C) contents, δ15N signatures, activities of N acquiring extracellular enzymes as well as the abundance of soil bacteria and fungi, and diazotrophs in bulk topsoil and rhizosphere were determined. Relative fungal abundance in the rhizosphere was higher under woodland and forest than under shrubland. This indicates toward plants' higher C investment into fungi in the Mediterranean and temperate rainforest sites than in the arid site. Fungi are likely to decompose lignified forest litter for efficient recycling of litter-derived N and further nutrients. Rhizosphere—a hotspot for the N fixation—was enriched in diazotrophs (factor 8 to 16 in comparison to bulk topsoil) emphasizing the general importance of root/microbe association in N cycle. These results show that the temperate rainforest is an N acquiring ecosystem, whereas N in the arid shrubland is strongly recycled. Simultaneously, the strongest 15N enrichment with decreasing N content with depth was detected in the Mediterranean woodland, indicating that N mineralization and loss is highest (and likely the fastest) in the woodland across the continental transect. Higher relative aminopeptidase activities in the woodland than in the forest enabled a fast N mineralization. Relative aminopeptidase activities were highest in the arid shrubland. The highest absolute chitinase activities were observed in the forest. This likely demonstrates that (a) plants and microorganisms in the arid shrubland invest largely into mobilization and reutilization of organically bound N by exoenzymes, and (b) that the ecosystem N nutrition shifts from a peptide-based N in the arid shrubland to a peptide- and chitin-based N nutrition in the temperate rainforest, where the high N demand is complemented by intensive N fixation in the rhizosphere. KW - Nitrogen KW - Rhizosphere KW - Microbial abundance KW - Natural abundance of 15N KW - Nitrogen fixation KW - Nitrogen uptake PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-543708 DO - https://doi.org/10.3389/fsoil.2022.817641 SN - 2673-8619 VL - 2 SP - 1 EP - 14 PB - Frontiers Media CY - Lausanne AN - OPUS4-54370 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Ruibal, C. A1 - Gueidan, C. A1 - Selbmann, L. A1 - Gorbushina, Anna A1 - Crous, P.W. A1 - Groenewald, J.Z. A1 - Muggia, L. A1 - Grube, M. A1 - Isola, D. A1 - Schoch, C.L. A1 - Staley, J.T. A1 - Lutzoni, F. A1 - De Hoog, G.S. T1 - Phylogeny of rock-inhabiting fungi related to Dothideomycetes N2 - The class Dothideomycetes (along with Eurotiomycetes) includes numerous rock-inhabiting fungi (RIF), a group of ascomycetes that tolerates surprisingly well harsh conditions prevailing on rock surfaces. Despite their convergent morphology and physiology, RIF are phylogenetically highly diverse in Dothideomycetes. However, the positions of main groups of RIF in this class remain unclear due to the lack of a strong phylogenetic framework. Moreover, connections between rock-dwelling habit and other lifestyles found in Dothideomycetes such as plant pathogens, saprobes and lichen-forming fungi are still unexplored. Based on multigene phylogenetic analyses, we report that RIF belong to Capnodiales (particularly to the family Teratosphaeriaceae s.l.), Dothideales, Pleosporales, and Myriangiales, as well as some uncharacterised groups with affinities to Dothideomycetes. Moreover, one lineage consisting exclusively of RIF proved to be closely related to Arthoniomycetes, the sister class of Dothideomycetes. The broad phylogenetic amplitude of RIF in Dothideomycetes suggests that total species richness in this class remains underestimated. Composition of some RIF-rich lineages suggests that rock surfaces are reservoirs for plant-associated fungi or saprobes, although other data also agree with rocks as a primary substrate for ancient fungal lineages. According to the current sampling, long distance dispersal seems to be common for RIF. Dothideomycetes lineages comprising lichens also include RIF, suggesting a possible link between rock-dwelling habit and lichenisation. KW - Arthoniomycetes KW - Capnodiales KW - Dothideomycetes KW - Evolution KW - Extremotolerance KW - Multigene phylogeny KW - Rock-inhabiting fungi PY - 2009 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-219821 DO - https://doi.org/10.3114/sim.2009.64.06 SN - 0166-0616 VL - 64 IS - 1 SP - 123 EP - 133 PB - Centraalbureau voor Schimmelcultures CY - Utrecht, The Netherlands AN - OPUS4-21982 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Gerrits, Ruben A1 - Pokharel, R. A1 - Breitenbach, Romy A1 - Radnik, Jörg A1 - Feldmann, Ines A1 - Schuessler, J.A. A1 - von Blanckenburg, F. A1 - Gorbushina, Anna A1 - Schott, J. T1 - How the rock-inhabiting fungus K. petricola A95 enhances olivine dissolution through attachment N2 - Free-living and mycorrhizal fungi are able to enhance the weathering of rock and other solid substrates. Deciphering the exact mechanisms of these natural processes requires their experimental simulation. Moreover, by performing these simulations with genetically amenable rock-weathering fungi, one can knock-out certain fungal traits and consequently identify their weathering-relevant function. Here, the effect of the rock-inhabiting fungus, Knufia petricola A95, on the dissolution kinetics of an Fe-bearing olivine (Mg1.86Fe0.19SiO4) is investigated at 25 °C and pH 6 using reproducible batch and mixed flow experiments. The availability of a melanin-deficient mutant (ΔKppks) of K. petricola A95, which produces more extracellular polymeric substances (EPS) than the wild type (WT), enables the comparative study of the role of melanin and EPS in olivine dissolution. In abiotic dissolution experiments, the olivine dissolution rate decreased considerably over time at pH 6 but not at pH 3.5. This inhibition of abiotic olivine dissolution at pH 6 was most likely caused by the in-situ oxidation of ferrous Fe and/or the precipitation of ferric hydroxides at the olivine surface. In corresponding biotic experiments at pH 6, both the wild type K. petricola and its melanin-deficient mutant ΔKppks solubilised and bound significant amounts of Fe released by olivine dissolution. Fe oxidation and precipitation were thus prevented and olivine dissolution proceeded faster than in the abiotic experiments. By sequestering Fe directly at the olivine surface, the attached wild type K. petricola cells were particularly efficient at preventing the oxidation of Fe at the mineral surface: the slowdown of olivine dissolution almost completely disappeared. The attachment capacity of these wild type cells is most likely mediated by wild type-specific EPS. Our presented experimental systems allow the oxidation of mineral-released Fe and include a rock-inhabiting fungus, thus simulating chemical, physical and biological conditions that set dissolution rates in a way that is relevant to natural ecosystems. KW - Black fungi KW - Bio-weathering KW - Forsterite KW - Knock-out mutant KW - Extracellular polymeric substances KW - Melanin Adhesion PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-509332 DO - https://doi.org/10.1016/j.gca.2020.05.010 VL - 282 SP - 76 EP - 97 PB - Elsevier Ltd. AN - OPUS4-50933 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Fuentes, Elsa A1 - Prieto, Beatriz A1 - Gorbushina, Anna T1 - Mixed model biofilm: interactions between algae, fungi and cyanobacteria and their effects on granite biodeterioration N2 - Granite, despite being one of the most widely used rocks for building material cultural heritage, is also one of the least studied. Previous works have shown that environmental parameters modulated by climate change will lead to changes in the microbial composition of biofilms grown on stone, but also to changes in the production of EPS, in the composition of pigments and, as a consequence, in the biodeterioration processes that this rock undergoes (Fuentes, 2023). Considering that the colonisation process is highly dynamic, small changes in climatic conditions can lead to the formation of different types of biofilms and diverse interactions between species. Knowing the contribution of the different organisms to the deterioration process is important both to anticipate the consequences that climate change will have on the deterioration of the granite, but also when addressing the processes of recolonization after cleaning, to ensure an improvement of the present situation. So, in order to continue my line of research on granite biodeterioration, the aim of this new project is to study the contribution of each type of organism (green algae, cyanobacteria and fungi) to the process of granite deterioration as well as the specific effect on each mineral, in addition to its role in the resilience and response of biofilms to environmental changes. With this main objective, the effect of the type of substrate on the composition of a model mixed biofilm formed by the fungus Knufia petricola, the algae Jaagichlorella sp. and the cyanobacterium Synechocystis sp. will be studied. The results obtained on granite will be compared with those obtained with other lithic types such as marble or limestone. Furthermore, it will be observed if the different ratios between organisms lead to changes in the capacity of these organisms to generate biodeterioration and to buffer climatic changes. Finally, the susceptibility of each mineral that makes up the granite - quartz, feldspars, micas - to biodeterioration mediated by each type of organism will be analysed. To address these questions, on the one hand, the response of the mixed biofilms as a whole will be evaluated by means of confocal microscopy, PAM, as well as the quantification of pigments and EPS, and on the other hand, the effect of these organisms on the deterioration of the substrates, through the use of SEM and TEM, which will allow the assessment of both the changes in porosity and the appearance of deterioration in the surface layers of the rock. This research will deepen the knowledge of the processes of granite deterioration, which have been limitedly studied in relation to microorganisms; but also aims to provide the scientific community with a model mixed biofilm to increase the reproducibility between experiments, one of the main limitations of this field of study. T2 - 19th International Biodeterioration and Biodegradation Symposium CY - Berlin, Germany DA - 09.09.2024 KW - Biofilm KW - Stone KW - Algae KW - Fungi KW - Cyanobacteria KW - Cultural heritage PY - 2024 AN - OPUS4-62272 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Seiffert, Franz A1 - Bandow, Nicole A1 - Kalbe, Ute A1 - Milke, R. A1 - Gorbushina, Anna T1 - Laboratory tools to quantify biogenic dissolution of rocks and minerals: a model rock biofilm growing in percolation columns N2 - Sub-aerial biofilms (SAB) are ubiquitous, self-sufficient microbial ecosystems found on mineral surfaces at all altitudes and latitudes. SABs, which are the principal causes of weathering on exposed terrestrial surfaces, are characterized by patchy growth dominated by associations of algae, cyanobacteria, fungi and heterotrophic bacteria. A recently developed in vitro system to study colonization of rocks exposed to air included two key SAB participants - the rock-inhabiting ascomycete Knufia petricola (CBS 123872) and the phototrophic cyanobacterium Nostoc punctiforme ATCC29133. Both partners are genetically tractable and we used them here to study weathering of granite, K-feldspar and plagioclase. Small fragments of the various rocks or minerals (1–6 mm) were packed into flow-through columns and incubated with 0.1% glucose and 10 μM thiamine-hydrochloride (90 μL min−1) to compare weathering with and without biofilms. Dissolution of the minerals was followed by: (i) analysing the degradation products in the effluent from the columns via Inductively Coupled Plasma Spectroscopy and (ii) by studying polished sections of the incubated mineral fragments/grains using scanning electron microscopy, transmission electron microscopy and energy dispersive X-ray analyses. K. petricola/N. punctiforme stimulated release of Ca, Na, Mg and Mn. Analyses of the polished sections confirmed depletion of Ca, Na and K near the surface of the fragments. The abrupt decrease in Ca concentration observed in peripheral areas of plagioclase fragments favored a dissolution-reprecipitation mechanism. Percolation columns in combination with a model biofilm can thus be used to study weathering in closed systems. Columns can easily be filled with different minerals and biofilms, the effluent as well as grains can be collected after long-term exposure under axenic conditions and easily analyzed. KW - Biotic weathering KW - Flow-through columns KW - Plagioclase KW - K-feldspar KW - Granite PY - 2016 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-373395 DO - https://doi.org/10.3389/feart.2016.00031 SN - 2296-6463 VL - 4 SP - Article 31, 1 EP - 10 PB - Frontiers in Earth Science CY - Lausanne AN - OPUS4-37339 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Lukowiak, M.C. A1 - Wettmarshausen, Sascha A1 - Hidde, Gundula A1 - Landsberger, Petra A1 - Boenke, Viola A1 - Rodenacker, K. A1 - Braun, Ulrike A1 - Friedrich, Jörg Florian A1 - Gorbushina, Anna A1 - Haag, R. T1 - Polyglycerol coated polypropylene surfaces for protein and bacteria resistance N2 - Polyglycerol (PG) coated polypropylene (PP) films were synthesized in a two-step approach that involved plasma bromination and subsequently grafting hyperbranched polyglycerols with very few amino functionalities. The influence of different molecular weights and density of reactive linkers were investigated for the grafted PGs. Longer bromination times and higher amounts of linkers on the surface afforded long-term stability. The protein adsorption and bacteria attachment of the PP-PG films were studied. Their extremely low amine content proved to be beneficial for preventing bacteria attachment. PY - 2015 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-325406 DO - https://doi.org/10.1039/c4py01375a SN - 1759-9954 SN - 1759-9962 VL - 6 IS - 8 SP - 1350 EP - 1359 AN - OPUS4-32540 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Menzel, Friederike A1 - Conradi, Bianca A1 - Rodenacker, K. A1 - Gorbushina, Anna A1 - Schwibbert, Karin ED - Menzel, Friederike T1 - Flow chamber system for the statistical evaluation of bacterial colonization on materials N2 - Biofilm formation on materials leads to high costs in industrial processes, as well as in medical applications. This fact has stimulated interest in the development of new materials with improved surfaces to reduce bacterial colonization. Standardized tests relying on statistical evidence are indispensable to evaluate the quality and safety of these new materials. We describe here a flow chamber system for biofilm cultivation under controlled conditions with a total capacity for testing up to 32 samples in parallel. In order to quantify the surface colonization, bacterial cells were DAPI (4‘,6-diamidino-2-phenylindole)-stained and examined with epifluorescence microscopy. More than 100 images of each sample were automatically taken and the surface coverage was estimated using the free open source software g’mic, followed by a precise statistical evaluation. Overview images of all gathered pictures were generated to dissect the colonization characteristics of the selected model organism Escherichia coli W3310 on different materials (glass and implant steel). With our approach, differences in bacterial colonization on different materials can be quantified in a statistically validated manner. This reliable test procedure will support the design of improved materials for medical, industrial, and environmental (subaquatic or subaerial) applications. KW - Subaerial and subaquatic biofilm KW - Escherichia coli KW - Image analysis KW - Microscopy KW - Biofilm KW - Biofouling PY - 2016 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-377040 DO - https://doi.org/10.3390/ma9090770 SN - 1996-1944 VL - 9 IS - 9 SP - 770 AN - OPUS4-37704 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 - Voigt, Oliver A1 - Knabe, Nicole A1 - Nitsche, Sarah A1 - Erdmann, Eileen A1 - Schumacher, Julia A1 - Gorbushina, Anna T1 - An advanced genetic toolkit for exploring the biology of the rock‑inhabiting black fungus Knufia petricola N2 - 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. KW - Subaerial biofilms KW - Biodeterioration KW - Fluorescent microscopy KW - CRISPR/Cas9 KW - RNA interference PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-518601 DO - https://doi.org/10.1038/s41598-020-79120-5 VL - 10 IS - 1 SP - 22021 PB - Springer Nature AN - OPUS4-51860 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Plarre, Rüdiger A1 - Zocca, Andrea A1 - Spitzer, Andrea A1 - Benemann, Sigrid A1 - Gorbushina, Anna A1 - Li, Y. A1 - Waske, Anja A1 - Funk, Alexander A1 - Wilgig, Janka A1 - Günster, Jens T1 - Searching for biological feedstock material: 3D printing of wood particles from house borer and drywood termite frass N2 - Frass (fine powdery refuse or fragile perforated wood produced by the activity of boring insects) of larvae of the European house borer (EHB) and of drywood termites was tested as a natural and novel feedstock for 3D-printing of wood-based materials. Small particles produced by the drywood termite Incisitermes marginipennis and the EHB Hylotrupes bajulus during feeding in construction timber, were used. Frass is a powdery material of particularly consistent quality that is essentially biologically processed wood mixed with debris of wood and faeces. The filigree-like particles flow easily permitting the build-up of woodbased structures in a layer wise fashion using the Binder Jetting printing process. The Quality of powders produced by different insect species was compared along with the processing steps and properties of the printed parts. Drywood termite frass with a Hausner Ratio HR = 1.1 with ρBulk = 0.67 g/cm3 and ρTap = 0.74 g/cm3 was perfectly suited to deposition of uniformly packed layers in 3D printing. We suggest that a variety of naturally available feedstocks could be used in environmentally responsible approaches to scientific material sciences/additive manufacturing. KW - 3D printing KW - X-ray tomographic KW - SEM micrography KW - Drywood termite PY - 2021 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-521517 DO - https://doi.org/10.1371/journal.pone.0246511 VL - 16 IS - 2 SP - e0246511 AN - OPUS4-52151 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Mitova, M. A1 - Iliev, M. A1 - Nováková, A. A1 - Gorbushina, Anna A1 - Groudeva, V.I. A1 - Martin-Sanchez, Pedro Maria T1 - Diversity and biocide susceptibility of fungal assemblages dwelling in the Art Gallery of Magura Cave, Bulgaria N2 - Magura Cave, north-western Bulgaria, possesses valuable rock-art paintings made with bat guano and dated from the period between the Eneolithic and Bronze Ages. Since 2008, the Art Gallery is closed to the general public in order to protect the paintings from vandalism, microclimatic changes caused by visitors and artificial illumination, and the consequent growth of fungi and phototrophs. Nevertheless, some tourist visits are allowed under the supervision of cave managers. This study provides the first scientific report on cultivable fungal assemblages dwelling different substrata in the Art Gallery. A total of 78 strains, belonging to 37 OTUs (Ascomycota 81%, Zygomycota 13%, Basidiomycota 5%), were isolated in the study. This fungal diversity was clearly dominated by Penicillium (50% of strains) and Aspergillus (13%). The most relevant visible fungal colonies were detected in sediments rich in bat guano, where, besides Penicillium, other guanophilic fungi such as Mucor, Mortierella, Trichosporon and Trichoderma were dominant. Conversely, scarce fungi were detected on rock surface of painted walls. Based on the biocide susceptibility assay, octylisothiazolinone (OIT) and benzalkonium chloride (BAC) were effective inhibiting the in vitro growth of dominant fungal species in Magura Cave, when applied at concentrations ranged from 100 to 1,000 mg/L. These data provide a valuable knowledge about Magura fungi, and exemplify a type of preliminary test that may be conducted before planning any biocide treatment. However, considering the irreversible effects of biocides on the ecological balance in caves, and the low fungal contamination in painted walls of Magura Cave, there is no reason to use conventional biocides in this cave. Further studies, monitoring microbial communities and microclimatic parameters, should be conducted to improve the knowledge on microbial ecology in Magura Cave and possible human impacts, as well as to allow the early detection of potential microbial outbreaks. KW - Fungi KW - Rock-art caves KW - Biocides PY - 2017 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-397436 DO - https://doi.org/10.5038/1827-806X.46.1.2061 SN - 0392-6672 VL - 46 IS - 1 SP - 67 EP - 80 PB - Scholar Commons CY - Tampa, FL, USA AN - OPUS4-39743 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Del Carmen Montero-Calasanz, M. A1 - Göker, M. A1 - Rohde, M. A1 - Schumann, P. A1 - Pötter, G. A1 - Spröer, C. A1 - Gorbushina, Anna A1 - Klenk, H.-P. T1 - Geodermatophilus siccatus sp. nov., isolated from arid sand of the Saharan desert in Chad N2 - A novel Gram-positive, aerobic, actinobacterial strain, CF6/1T, was isolated in 2007 during environmental screening of arid desert soil in the Sahara near to Ourba, Chad. The isolate was found to grow best in a temperature range of 20–37 °C and at pH 6.0–8.5 and showed no NaCl tolerance, forming black-coloured and nearly circular colonies on GYM agar. Chemotaxonomic and molecular characteristics determined for the isolate match those previously described for members of the genus Geodermatophilus. The DNA G + C content of the novel strain was determined to be 74.9 mol %. The peptidoglycan was found to contain meso-diaminopimelic acid as the diagnostic diamino acid. The main phospholipids were determined to be phosphatidylethanolamine, phosphatidylinositol, phosphatidylcholine, diphosphatidylglycerol and traces of phosphatidylglycerol; MK-9(H4) was identified as the dominant menaquinone and galactose as the diagnostic sugar. The major cellular fatty acids were found to be the branched-chain saturated acids iso-C16:0 and iso-C15:0, as well as C17:1ω8c. The 16S rRNA gene sequence shows 97.5–97.9 % sequence identity with the four validly named or at least effectively published members of the genus: Geodermatophilus obscurus (97.5 %), Geodermatophilus arenarius (97.7 %), Geodermatophilus ruber (97.9 %) and Geodermatophilus nigrescens (97.9 %). Based on the results from this polyphasic taxonomic analysis and DNA–DNA hybridizations with all type strains of the genus, we propose that strain CF6/1T represents a novel species, Geodermatophilus siccatus, with the type strain CF6/1T = DSM 45419T = CCUG 62765T = MTCC 11414T. KW - Actinomycetes KW - Geodermatophilaceae KW - Taxonomy KW - Xerophiles KW - Phenotype microarray PY - 2013 DO - https://doi.org/10.1007/s10482-012-9824-x SN - 0003-6072 SN - 1572-9699 VL - 103 IS - 3 SP - 449 EP - 456 PB - Springer CY - Dordrecht AN - OPUS4-28082 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Microbiology / biofilms in material research and testing N2 - In the modern world there is an increased understanding that design and performance monitoring of materials have to be tested in connection to chemical, physical and (micro)biological challenges. A systematic study on how biofilms interact with materials and what could be done to engineer biofilms and/or materials in order to maximize the resistance of the material (surface) or the resistance the biofilm-modified material (bulk) is in strong need. In the Department “Materials and the Environment” of the BAM new experimental platform is being developed. With the help of different type of device for high throughput and microbiologically-controlled environment simulation we establish a new approach to clarify the mechanisms of biofilm/material interactions. Despite the focus on fundamental research, the main results of this project proposal will be transferable into material technology and construction chemistry and will influence the development of standardization in this topic. As the interactions of biofilms and materials have implications for most constructions as well as climate change, the results of the research generates additional value. T2 - Initialgespräch - DFG-Forschungsgruppe "Mikrobiologie/Biofilme" CY - Karlsruhe, Germany DA - 14.11.2019 KW - Biofilm KW - Microbiology KW - Black fungi KW - Solar panel PY - 2019 AN - OPUS4-50199 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Breitenbach, Romy A1 - Gerrits, Ruben A1 - Dementyeva, Polina A1 - Knabe, Nicole A1 - Schumacher, Julia A1 - Feldmann, Ines A1 - Radnik, Jörg A1 - Ryo, M. A1 - Gorbushina, Anna T1 - The role of extracellular polymeric substances of fungal biofilms in mineral attachment and weathering N2 - The roles extracellular polymeric substances (EPS) play in mineral attachment and weathering were studied using genetically modified biofilms of the rock-inhabiting fungus Knufia petricola strain A95. Mutants deficient in melanin and/or carotenoid synthesis were grown as air-exposed biofilms. Extracted EPS were quantified and characterised using a combination of analytical techniques. The absence of melanin affected the quantity and composition of the produced EPS: mutants no longer able to form melanin synthesised more EPS containing fewer pullulan-related glycosidic linkages. Moreover, the melanin-producing strains attached more strongly to the mineral olivine and dissolved it at a higher rate. We hypothesise that the pullulan-related linkages, with their known adhesion functionality, enable fungal attachment and weathering. The released phenolic intermediates of melanin synthesis in the Δsdh1 mutant might play a role similar to Fe-chelating siderophores, driving olivine dissolution even further. These data demonstrate the need for careful compositional and quantitative analyses of biofilm-created microenvironments. KW - Biofilms PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-549025 DO - https://doi.org/10.1038/s41529-022-00253-1 SN - 2397-2106 VL - 6 SP - 1 EP - 11 PB - Springer Nature CY - London AN - OPUS4-54902 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Alder-Rangel, A. A1 - Idnurm, A. A1 - Brand, A. A1 - Brown, A. A1 - Gorbushina, Anna A1 - Kelliher, C. A1 - Campos, C. A1 - Levin, D. A1 - Bell-Pedersen, D. A1 - Dadachova, E. A1 - Bauer, F. A1 - Gadd, G. A1 - Braus, G. A1 - Braga, G. A1 - Brancini, G. A1 - Walker, G. A1 - Druzhinina, I. A1 - Pocsi, I. A1 - Dijksterhuis, J. A1 - Aguirre, J. A1 - Hallsworth, J. A1 - Schumacher, Julia A1 - Ho Wong, K. A1 - Selbmann, L. A1 - Corrochano, L. A1 - Kupiec, M. A1 - Momany, M. A1 - Molin, M. A1 - Requena, N. A1 - Yarden, O. A1 - Cordero, R. A1 - Fischer, R. A1 - Pascon, R. A1 - Mancinelli, R. A1 - Emri, T. A1 - Basso, T. A1 - Rangel, D. T1 - The Third International Symposium on Fungal Stress - ISFUS N2 - Stress is a normal part of life for fungi, which can survive in environments considered inhospitable or hostile for other organisms. Due to the ability of fungi to respond to, survive in, and transform the environment, even under severe stresses, many researchers are exploring the mechanisms that enable fungi to adapt to stress. The International Symposium on Fungal Stress (ISFUS) brings together leading scientists from around the world who research fungal stress. This article discusses presentations given at the third ISFUS, held in Sao Jose dos Campos, Sao Paulo, Brazil in 2019, thereby summarizing the state-of-the-art knowledge on fungal stress, a field that includes microbiology, agriculture, ecology, biotechnology, medicine, and astrobiology. T2 - International Symposium on Fungal Stress (ISFUS) CY - Sao Jose dos Campos, Brazil DA - 20.05.2019 KW - Agricultural mycology KW - Fungal stress mechanisms and responses KW - Industrial mycology KW - Medical mycology PY - 2020 DO - https://doi.org/10.1016/j.funbio.2020.02.007 VL - 124 IS - 5 SP - 235 EP - 252 PB - Elsevier Ltd. AN - OPUS4-50953 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Favet, J. A1 - Lapanje, A. A1 - Giongo, A. A1 - Kennedy, S. A1 - Aung, Y.-Y. A1 - Cattaneo, A. A1 - Davis-Richardson, A.G. A1 - Brown, C.T. A1 - Kort, R. A1 - Brumsack, H.-J. A1 - Schnetger, B. A1 - Chappell, A. A1 - Kroijenga, J. A1 - Beck, A. A1 - Schwibbert, Karin A1 - Mohamed, A.H. A1 - Kirchner, T. A1 - Dorr de Quadros, P. A1 - Triplett, E.W. A1 - Broughton, William J. A1 - Gorbushina, Anna T1 - Microbial hitchhikers on intercontinental dust: catching a lift in Chad N2 - Ancient mariners knew that dust whipped up from deserts by strong winds travelled long distances, including over oceans. Satellite remote sensing revealed major dust sources across the Sahara. Indeed, the Bodélé Depression in the Republic of Chad has been called the dustiest place on earth. We analysed desert sand from various locations in Chad and dust that had blown to the Cape Verde Islands. High throughput sequencing techniques combined with classical microbiological methods showed that the samples contained a large variety of microbes well adapted to the harsh desert conditions. The most abundant bacterial groupings in four different phyla included: (a) Firmicutes—Bacillaceae, (b) Actinobacteria—Geodermatophilaceae, Nocardiodaceae and Solirubrobacteraceae, (c) Proteobacteria—Oxalobacteraceae, Rhizobiales and Sphingomonadaceae, and (d) Bacteroidetes—Cytophagaceae. Ascomycota was the overwhelmingly dominant fungal group followed by Basidiomycota and traces of Chytridiomycota, Microsporidia and Glomeromycota. Two freshwater algae (Trebouxiophyceae) were isolated. Most predominant taxa are widely distributed land inhabitants that are common in soil and on the surfaces of plants. Examples include Bradyrhizobium spp. that nodulate and fix nitrogen in Acacia species, the predominant trees of the Sahara as well as Herbaspirillum (Oxalobacteraceae), a group of chemoorganotrophic free-living soil inhabitants that fix nitrogen in association with Gramineae roots. Few pathogenic strains were found, suggesting that African dust is not a large threat to public health. KW - Aeolian KW - High throughput sequencing KW - Bodélé Depression KW - Republic of Chad KW - Wind erosion PY - 2013 DO - https://doi.org/10.1038/ismej.2012.152 SN - 1751-7362 SN - 1751-7370 VL - 7 SP - 850 EP - 867 PB - Nature Publishing Group CY - Basingstoke AN - OPUS4-28083 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Vogel, Christian A1 - Urban, Klaus A1 - Ackerhans, C. A1 - Gorbushina, Anna T1 - Analysis of Carbon and Nitrogen from Atmospheric Sources by Bulk Deposition Sampling at various locations in Germany N2 - Atmospheric deposition of particulate matter is an important indicator of air pollution and a significant factor in material surface fouling. The elemental composition of this nutrient-containing dust depends largely on the exposure region and time as well as on climate. Therefore, in this paper we report an analysis of atmospheric pollutions with a self-made low-cost bulk deposition sampler directed at sampling deposition via air transport and rainfall. We used the device in diverse environments - thus comparing an urban region, an area surrounded by forest and an area mainly dominated by agriculture. The total organic carbon (TOC) and total nitrogen (TN) amounts were selected as indicator parameters and analyzed in a biweekly rhythm for three and a half and two years, respectively. The TOC value responded to particulate matter in the urban area, especially significant were the influences of the New Year's firework in urban and pollen in the rural forest area. In contrast, the TN value was more under the influence of the nitrogen emissions in the agriculture-dominated area. However, the TN value did not correlate with the NOx values in the urban area because the atmospheric nitrogen emissions in the city might originate from various emission sources. Summarizing, the TOC and TN values of the self-made low-cost bulk deposition sampler were in good agreement with environmental events of their immediate surrounding. Moreover, the selected containers and sampling procedures are universally applicable to monitor and analyze organic as well as inorganic parameters (e.g. metal ions) of atmospheric deposition. KW - Passive sampling KW - Biomonitoring KW - Air Analysis PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-609373 DO - https://doi.org/10.1016/j.envadv.2024.100583 SN - 2666-7657 VL - 17 SP - 1 EP - 9 PB - Elsevier Ltd. AN - OPUS4-60937 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Gesteinsbesiedelnde Pilze als Modellorganismen T2 - Vortrag CY - Rostock, Germany DA - 2011-04-14 PY - 2011 AN - OPUS4-23560 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Stock, S. C. A1 - Koester, M. A1 - Boy, J. A1 - Godoy, R. A1 - Nájera, F. A1 - Matus, F.J. A1 - Merino, C. A1 - Abdallah, Khaled A1 - Leuschner, C. A1 - Spielvogel, S. A1 - Gorbushina, Anna A1 - Kuzyakov, Y. A1 - Dippold, M. A. T1 - Plant carbon investment infine roots and arbuscular mycorrhizal fungi: Across-biome study on nutrient acquisition strategies N2 - Plant resource acquisition strategies were ecosystem-specific with distinct mutualism with arbuscular mycorrhizal (AM) fungi. Root traits indicated conservative resource economics in the arid shrubland, but an acquisitive and self-sufficient (“do-it-yourself”) acquisition strategy in the semiarid coastal matorral, resulting in large carbon (C) investments (green). Forest plants with conservative root traits seem to intensively outsource their acquisition to AM fungi, compensating for lower uptake capacities of conservative roots (red line). High allocations of freshly assimilated C into AM fungal storage compounds illustrated the relevance of AM fungi as C sink, especially in the semiarid matorral. KW - Natural ecosystems KW - Temperate rain forest KW - Arbuscular mycorrhiza KW - Plant economic spectrum KW - Root economics space KW - 13CO2 pulse labeling PY - 2021 DO - https://doi.org/10.1016/j.scitotenv.2021.146748 VL - 781 SP - 146748 PB - Elsevier B.V. CY - Amsterdam AN - OPUS4-52804 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Selbmann, L. A1 - Benkő, Z. A1 - Coleine, C. A1 - de Hoog, S. A1 - Donati, C. A1 - Druzhinina, I. A1 - Emri, T. A1 - Ettinger, C. L. A1 - Gladfelter, A. S. A1 - Gorbushina, Anna A1 - Grigoriev, I. V. A1 - Grube, M. A1 - Gunde-Cimerman, N. A1 - Karányi, Z. A. A1 - Kocsis, B. A1 - Kubressoian, T. A1 - Miklós, I. A1 - Miskei, M. A1 - Muggia, L. A1 - Northen, T. A1 - Novak-Babič, M. A1 - Pennacchio, C. A1 - Pfliegler, W. P. A1 - Pòcsi, I. A1 - Prigione, V. A1 - Riquelme, M. A1 - Segata, N. A1 - Schumacher, Julia A1 - Shelest, E. A1 - Sterflinger, K. A1 - Tesei, D. A1 - U’Ren, J. M. A1 - Varese, G. C. A1 - Vázquez-Campos, X. A1 - Vicente, V. A. A1 - Souza, E. M. A1 - Zalar, P. A1 - Walker, A. K. A1 - Stajich, J. E. T1 - Shed Light in the DaRk LineagES of the Fungal Tree of Life—STRES N2 - The polyphyletic group of black fungi within the Ascomycota (Arthoniomycetes, Dothideomycetes, and Eurotiomycetes) is ubiquitous in natural and anthropogenic habitats. Partly because of their dark, melanin-based pigmentation, black fungi are resistant to stresses including UV- and ionizing-radiation, heat and desiccation, toxic metals, and organic pollutants. Consequently, they are amongst the most stunning extremophiles and poly-extreme-tolerant organisms on Earth. Even though ca. 60 black fungal genomes have been sequenced to date, [mostly in the family Herpotrichiellaceae (Eurotiomycetes)], the class Dothideomycetes that hosts the largest majority of extremophiles has only been sparsely sampled. By sequencing up to 92 species that will become reference genomes, the “Shed light in The daRk lineagES of the fungal tree of life” (STRES) project will cover a broad collection of black fungal diversity spread throughout the Fungal Tree of Life. Interestingly, the STRES project will focus on mostly unsampled genera that display different ecologies and life-styles (e.g., ant- and lichen-associated fungi, rock-inhabiting fungi, etc.). With a resequencing strategy of 10- to 15-fold depth coverage of up to ~550 strains, numerous new reference genomes will be established. To identify metabolites and functional processes, these new genomic resources will be enriched with metabolomics analyses coupled with transcriptomics experiments on selected species under various stress conditions (salinity, dryness, UV radiation, oligotrophy). The data acquired will serve as a reference and foundation for establishing an encyclopedic database for fungal metagenomics as well as the biology, evolution, and ecology of the fungi in extreme environments. KW - Adaptation KW - Black fungi KW - Dothideomycetes KW - Eurotiomycetes KW - Extremophiles KW - Genomics KW - Metabolomics KW - Secondary metabolites KW - Stress conditions KW - Transcriptomics PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-519328 DO - https://doi.org/10.3390/life10120362 VL - 10 IS - 12 SP - 362 PB - MDPI CY - Basel AN - OPUS4-51932 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CHAP A1 - Lessovaia, S.N. A1 - Gerrits, Ruben A1 - Gorbushina, Anna A1 - Polekhovsky, Y.S. A1 - Dultz, S. A1 - Kopitsa, G.G. ED - Frank-Kamenetskaya, O.V. ED - Vlasov, D. ED - Panova, E.G. ED - Lessovaia, S.N. T1 - Modeling Biogenic Weathering of Rocks from Soils of Cold Environments N2 - Morphologically simple and microbially dominated ecosystems termed “biofilms” have existed on Earth for a long period of biosphere evolution. A model biofilm combining one heterotroph and one phototroph component was used in a laboratory experiment to simulate biogenic weathering with two different specimens of basic rock samples from the soil profiles. The rocks fragments from the regions of cold environments of Eurasia,where abiotic physical processes, including rock disintegration initiated by freezing–thawing cycles, represent the most probable Scenario of rock weathering, were subjected to biological colonization. The rock Fragments were represented by dolerite and metagabbro amphibolites. Polished sections of the rock samples were inoculated with the model microbiological consortium of the oligotrophic fungus and the phototrophic cyanobacteria (biofilm). After 3 month runtime of the experiment the progress of rock weathering was derived from the growth of the biofilm on the rock surfaces. The model biofilm visualization on the rock surface of polished sections illustrated their stronger development namely on dolerite in comparison with metagabbro amphibolite. The findings confirmed the higher sensitivity of dolerite to biogenic weathering due to (i) mineral association, in which quartz was absent and (ii) porosity providing higher specific surface area for biotic—abiotic interaction influenced by the occurrence of micro-porosity in the rock. KW - Biogenic weathering KW - Rock leaching KW - Fractal structure KW - Biofilm formation KW - Internal pores PY - 2020 DO - https://doi.org/10.1007/978-3-030-21614-6_27 SP - 501 EP - 515 PB - Springer AN - OPUS4-51442 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - A Genetic Toolbox for Exploring the Life Style of the Rock-inhabiting Black Fungus Knufia petricola N2 - After their discovery on rock surfaces in cold and hot deserts, a polyphyletic group of ascomycetous black fungi was found to dominate a range of hostile environments – natural and man-made, from salterns to dishwashers, roofs and solar panels. Together with bacteria and algae they may establish sub-aerial biofilms and cause weathering of the surfaces they grow on. Their impressive survival abilities as well as their constitutive protective pigmentation and cluster-like microcolony organisation are similar in environmental isolates as well as in heat-tolerant opportunistic pathogens of animals and humans. The exact genetic properties that ensure their survival in extreme environments can be studied if some black fungi were amenable to genetic manipulations. We selected the rock-inhabiting fungus Knufia petricola (class Eurotiomycetes, order Chaetothyriales) that grows moderately in axenic culture and exhibits all the characteristics of black yeasts such as yeast-like cell growth, absence of reproductive structures and constitutive dihydroxynaphthalene (DHN) melanogenesis. For this environmental strain we developed protocols to efficiently generate and transform protoplasts resulting in stable homokaryotic transformants by targeting genes involved in pigment synthesis and expressing fluorescent reporter genes. Hence, endogenous and foreign genes can be expressed from episomal AMA1-containing plasmids and genome-integrated DNA constructs. Moderate rates of homologous recombination allow for both ectopic and targeted integrations. CRISPR-Cas9 was further validated as a strategy for obtaining selection marker-free mutants and silencing via RNA interference as an approach to study essential genes. Availability of this genetic toolbox and an annotated genome sequence is paving the way for studying interactions of K. petricola and other black yeasts with environmental stressors, material surfaces, soil matrices and phototrophic symbionts. T2 - VAAM symposium 'Molecular Biology of Fungi' CY - Göttingen, Germany DA - 19.09.2019 KW - Knufia petricola KW - Rock-inhabiting fungus KW - Genetics KW - Crispr-Cas9 PY - 2019 AN - OPUS4-49634 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Stohl, Leonie A1 - Tonon, Chiara A1 - Cook, Jake A1 - Gorbushina, Anna A1 - Dehn, Frank A1 - von Werder, Julia A1 - Manninger, Tanja T1 - Understanding bioreceptivity of concrete: material design and characterization N2 - The climate crisis is driving an increasing demand for ecologically oriented concepts. In the building sector, this demand includes not only the use of environmentally friendly materials but also the greening of urban areas. One promising approach is the development of bioreceptive concrete façades, which support the growth of green biofilms directly on their surfaces. These innovative façades are anticipated to deliver benefits comparable to those of macroscopically greened façades, such as enhanced biodiversity and improved air quality, while offering the advantages of being more self-sustaining and stable systems once fully established. However, the development of bioreceptive concrete presents substantial challenges. Due to the interdisciplinarity and novelty of this field, standardized methods for material characterization and bioreceptivity assessment are currently lacking. This study proposes an approach for evaluating surface properties crucial for bioreceptivity, developed on differently structured samples of ultra-high-performance concrete (UHPC). Existing methods and standards from concrete technology are critically reviewed and, where necessary, modified to meet the unique requirements of measuring bioreceptive material properties. Special attention is given to the surface pH value and water retention characteristics, as these are essential for promoting microbial growth and ensuring the long-term stability of green biofilms. The observed surface characteristics vary according to the imprinted surface structures, offering a spectrum of material properties and enabling the evaluation of their impact on bioreceptivity. The findings presented form the foundation for subsequent laboratory weathering experiments, which will be discussed in a complementary publication. KW - Novel methods KW - Bioreceptivity testing PY - 2025 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-655023 DO - https://doi.org/10.1617/s11527-025-02863-y SN - 1359-5997 VL - 58 SP - 1 EP - 18 PB - Springer Science and Business Media LLC AN - OPUS4-65502 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CHAP A1 - Breitenbach, Romy A1 - Toepel, Jörg A1 - Dementyeva, Polina A1 - Knabe, Nicole A1 - Gorbushina, Anna ED - Flemming, H.-C. ED - Neu, T. R. ED - Wingener, J. T1 - Snapshots of fungal extracellular matrices N2 - Fungal extracellular materials reinforce a constant interaction between their cell wall and the environment. A dynamic mixture of chitin, glucans, mannans, glycoproteins, glycolipids and pigments supports the success of all fungal life styles – from symbiotic to the free-living and pathogenic. Fungi are perfectly adapted to grow on surfaces and in porous environments, where they form medically and geochemically relevant biofilms. Fungal EPS are critical in adhesion to other fungi, other cells or substratum as well as in the following interaction with the host immune system or material they attack, degrade and deteriorate respectively. Characterisation of extracellular compounds and understanding of its function is necessary to limit damage caused by fungal activity. All necessary methodology from chemical characterization to complete genetic analyses has been developed for medically important fungi. Now it is time to apply this knowledge to the numerous, largely aerobic and very active organisms that occupy a wide range of atmosphere-exposed habitats in the upper lithosphere. One can expect that analogies between medically- and environmentally-relevant model fungal species will help us to address the dynamics of the fungal cell EPS matrix in much more efficient and widely applicable ways. KW - Fungal cell wall KW - Melanin KW - Fungal life styles KW - Environmental and pathogenic fungi KW - Model fungal biofilms KW - Biogenic weathering PY - 2016 SN - 9781780407418 SN - 9781780407425 DO - https://doi.org/10.2166/9781780407418 SP - Chapter 14, 269 EP - 299 PB - IWA Publishing CY - London, UK AN - OPUS4-38094 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - From rocks to riches: Knufia petricola as model and expression platform N2 - Black fungi belong to different classes of Ascomycota but evolved similar morpho-physiological adaptations such as yeast-like growth and constitutive melanin formation to colonize extreme competition-free environments. They are ubiquitously found on air-exposed surfaces, from ancient marble monuments to modern solar panels. The rock inhabitant Knufia petricola was chosen to become a model for these extremotolerant black fungi. Plasmid-based and ribonucleoprotein-based CRISPR/Cas9 techniques were introduced to precisely introduce one to multiple double strand breaks into the DNA to modify, replace or add sequences to the genome either using the available selection marker systems (hygR, natR, genR, baR, suR) or by marker-free approaches. Multiplexing is very efficient, allowing for four or more simultaneous genome editing events. The newly generated cloning vectors containing the Tet on construct for doxycycline-controlled gene expression, and the validated sites in the K. petricola genome for color-selectable (pks1, phs1, ade2) or neutral insertion (igr1 to 5) of expression constructs complete the reverse genetics toolbox. One or multiple endogenous or heterologous genes can be expressed on demand from different genomic loci or from a single construct by using 2A self-cleaving peptides. We thus can express genes from synthetic clusters in a non-pigmented background (Δpks1/Δphs1). The fact that K. petricola only produces few secondary metabolites (DHN melanin, carotenoids, mycosporines and a siderophore) and plant cell-wall degrading enzymes but is capable to take the burden of acetyl-CoA-consuming metabolism and protein secretion renders K. petricola a promising host for the expression of heterologous genes encoding high-end secondary metabolites and enzymes. T2 - 12th International Mycological Congress (IMC12) CY - Maastricht, Netherlands DA - 11.08.2024 KW - Fungus KW - Genetic engineering KW - Heterologous gene expression KW - Cell factory PY - 2024 AN - OPUS4-60838 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - GEN A1 - Breitenbach, Romy A1 - Gerrits, Ruben A1 - Dementyeva, Polina A1 - Knabe, Nicole A1 - Schumacher, Julia A1 - Feldmann, Ines A1 - Radnik, Jörg A1 - Ryo, M. A1 - Gorbushina, Anna T1 - Data for "The role of extracellular polymeric substances of fungal biofilms in mineral attachment and weathering" N2 - Data for the publication "The role of extracellular polymeric substances of fungal biofilms in mineral attachment and weathering" (https://doi.org/10.1038/s41529-022-00253-1). It includes: - The Summary of the EPS concentration, EPS sugar components and EPS linkages. - The Summary of the XPS analysis of freeze-dried biofilm samples of all strains. - The Summary of the pH, Mg, SI and Fe concentration, biomass and olivine dissolution rate for each time point of all dissolution experiments. KW - Biofilms PY - 2022 DO - https://doi.org/10.26272/opus4-54901 PB - Bundesanstalt für Materialforschung und -prüfung (BAM) CY - Berlin AN - OPUS4-54901 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Wen, Keqing A1 - Gorbushina, Anna A1 - Schwibbert, Karin A1 - Bell, Jérémy T1 - Microfluidic platform with precisely controlled hydrodynamic parameters and integrated features for generation of microvortices to accurately form and monitor biofilms in flow N2 - Microorganisms often live in habitats characterized by fluid flow, and their adhesion to surfaces in industrial systems or clinical settings may lead to pipe clogging, microbially influenced corrosion, material deterioration, food spoilage, infections, and human illness. Here, a novel microfluidic platform was developed to investigate biofilm formation under precisely controlled (i) cell concentration, (ii) temperature, and (iii) flow conditions. The developed platform central unit is a single-channel microfluidic flow cell designed to ensure ultrahomogeneous flow and condition in its central area, where features, e.g., with trapping properties, can be incorporated. In comparison to static and macroflow chamber assays for biofilm studies, microfluidic chips allow in situ monitoring of biofilm formation under various flow regimes and have better environment control and smaller sample requirements. Flow simulations and experiments with fluorescent particles were used to simulate bacteria flow in the platform cell for calculating flow velocity and direction at the microscale level. The combination of flow analysis and fluorescent strain injection in the cell showed that microtraps placed at the center of the channel were efficient in capturing bacteria at determined positions and to study how flow conditions, especially microvortices, can affect biofilm formation. The microfluidic platform exhibited improved performances in terms of homogeneity and robustness for in vitro biofilm formation. We anticipate the presented platform to be suitable for broad, versatile, and high-throughput biofilm studies at the microscale level. KW - Topographical pattern KW - E. coli KW - Fluorescence KW - Bacteria trapping KW - Particle velocimetry PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-610450 DO - https://doi.org/10.1021/acsbiomaterials.4c00101 SN - 2373-9878 VL - 10 IS - 7 SP - 4626 EP - 4634 PB - ACS Publ. CY - Washington, DC AN - OPUS4-61045 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - A genetic toolbox for exploring the life style of the rock-inhabiting black fungus Knufia petricola N2 - After their discovery on rock surfaces in cold and hot deserts, a polyphyletic group of ascomycetous black fungi was found to dominate a range of hostile environments – natural and man-made, from salterns to dishwashers, roofs and solar panels. Together with bacteria and algae they may establish subaerial biofilms and cause weathering of the surfaces they grow on. Their impressive survival abilities as well as their constitutive protective pigmentation and cluster-like microcolony organisation are similar in environmental isolates as well as in heat-tolerant opportunistic pathogens of animals and humans. The exact genetic properties that ensure their survival in extreme environments can be studied if some black fungi were amenable to genetic manipulations. We selected the rock-inhabiting fungus Knufia petricola (class Eurotiomycetes, order Chaetothyriales) that grows moderately in axenic culture and exhibits all the characteristics of microcolonial black fungi such as yeast-like cell growth, absence of reproductive structures and constitutive dihydroxynaphthalene (DHN) melanogenesis (Nai et al. 2013, Fungal Genet Biol). We developed protocols to efficiently generate and transform protoplasts resulting in stable homokaryotic transformants by targeting genes involved in pigment synthesis and expressing fluorescent reporter genes. Hence, endogenous and foreign genes can be expressed from episomal AMA1-containing plasmids and genome-integrated DNA constructs. Moderate rates of homologous recombination allow for both ectopic and targeted integrations. CRISPR-Cas9 was further validated as a strategy for obtaining selection marker-free mutants and silencing via RNA interference as an approach to study essential genes. Availability of this genetic toolbox and an annotated genome sequence of the strain A95 is paving the way for studying interactions of K. petricola with environmental stressors, material surfaces, soil matrices and phototrophic symbionts. T2 - European Conference on Fungal Genetics (ECFG15) CY - Rome, Italy DA - 17.02.2020 KW - black fungi KW - genomics KW - genetics PY - 2020 AN - OPUS4-50593 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Catanzaro, Ilaria A1 - Gorbushina, Anna A1 - Onofri, Silvano A1 - Schumacher, Julia T1 - 1,8‐Dihydroxynaphthalene (DHN) melanin provides unequal protection to black fungi Knufia petricola and Cryomyces antarcticus from UV‐B radiation N2 - Black fungi on rock surfaces endure a spectrum of abiotic stresses, including UV radiation. Their ability to tolerate extreme conditions is attributed to the convergent evolution of adaptive traits, primarily highly melanized cell walls. However, studies on fungal melanins have not provided univocal results on their photoprotective functions. Here, we investigated whether the black fungi Knufia petricola and Cryomyces antarcticus only use DHN melanin or may employ alternative mechanisms to counteract UV‐induced damage. For this, melanized wild types and non‐melanized Δpks1 mutants were exposed to different doses of UV‐B (312 nm) followed by incubation in constant darkness or in light–dark cycles to allow light‐dependent DNA repair by photolyases (photoreactivation). C. antarcticus could tolerate higher UV‐B doses but was sensitive to white light, whereas K. petricola showed the opposite trend. DHN melanin provided UV‐B protection in C. antarcticus, whereas the same pigment or even carotenoids proved ineffective in K. petricola. Both fungi demonstrated functional photoreactivation in agreement with the presence of photolyase‐encoding genes. Our findings reveal that although the adaptive trait of DHN melanization commonly occurs across black fungi, it is not equally functional and that there are species‐specific adaptations towards either UV‐induced lesion avoidance or repair strategies. KW - Rock-inhabiting Ascomycetes KW - Pigment KW - Photoprotection KW - Photolyase KW - Disinfection PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-616859 DO - https://doi.org/10.1111/1758-2229.70043 VL - 16 IS - 6 SP - 1 EP - 13 PB - Wiley AN - OPUS4-61685 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Tan, K. H. A1 - Sattari, S. A1 - Donskyi, Ievgen A1 - Cuellar-Camacho, J. L. A1 - Cheng, C. A1 - Schwibbert, Karin A1 - Lippitz, Andreas A1 - Unger, Wolfgang A1 - Gorbushina, Anna A1 - Adeli, M. A1 - Haag, R. T1 - Functionalized 2D nanomaterials with switchable binding to investigate graphene–bacteria interactions N2 - Graphene and its derivatives have recently attracted much attention for sensing and deactivating pathogens. However, the mechanism of multivalent interactions at the graphene–pathogen interface is not fully understood. Since different physicochemical parameters of graphene play a role at this interface, control over graphene’s structure is necessary to study the mechanism of these interactions. In this work, different graphene derivatives and also zwitterionic graphene nanomaterials (ZGNMs) were synthesized with defined exposure, in terms of polymer coverage and functionality, and isoelectric points. Then, the switchable interactions of these nanomaterials with E. coli and Bacillus cereus were investigated to study the validity of the generally proposed “trapping” and “nano-knives” mechanisms for inactivating bacteria by graphene derivatives. It was found that the antibacterial activity of graphene derivatives strongly depends on the accessible area, i.e. edges and basal plane of sheets and tightness of their agglomerations. Our data clearly confirm the authenticity of “trapping” and “nano-knives” mechanisms for the antibacterial activity of graphene sheets. KW - XPS KW - Graphene KW - Graphene–bacteria interaction PY - 2018 DO - https://doi.org/10.1039/c8nr01347k SN - 2040-3364 SN - 2040-3372 VL - 10 IS - 20 SP - 9525 EP - 9537 PB - RSC CY - London AN - OPUS4-45084 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Gorbushina, Anna T1 - Nutritional physiology of Sarcinomyces petricola A95, a model black fungus to study primary successions in terrestrial ecosystems T2 - Fourth meeting of the ISHAM working groups on Black Yeasts and chromoblastomycosis: “Hidden Danger, Bright Promise” CY - Curitiba, Brazil DA - 2011-12-01 PY - 2011 AN - OPUS4-26470 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -