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 -