TY - CONF A1 - Schumacher, Julia A1 - Brandhorst, Antonia A1 - Catanzaro, Ilaria A1 - Nitsche, Sarah A1 - Erdmann, Eileen A1 - Gerrits, Ruben A1 - Gorbushina, Anna T1 - Versatile DHN melanin – spotlight on its function in microcolonial black fungi N2 - Dihydroxynaphthalene (DHN) melanin is produced by diverse Ascomycetes via slightly differing biosynthetic routes. The polyketide synthases (PKS) release the heptaketide YWA1, the hexaketide AT4HN or the pentaketide T4HN. The first two products are deacetylated by ‘yellowish-green’ hydrolases to T4HN, and T4HN is further converted by a core set of enzymes to DHN. Final polymerization steps are accomplished by multicopper oxidases. DHN melanogenesis is often regulated in a spatial and temporal fashion resulting e.g., in melanized reproduction and survival structures of the foliar plant pathogen Botrytis cinerea (Schumacher 2016, Mol Microbiol). In contrast, a polyphyletic group of Ascomycetes (microcolonial fungi/ black yeasts) dwelling in hostile habitats such as bare rock surfaces in hot and cold deserts, exhibits constitutive melanogenesis. Here, DHN melanin builds a protective layer around all vegetative cells thus contributing to the survival of diverse environmental stresses even without specialized reproduction structures. For studying the relevance of constitutive DHN melanogenesis for tolerance of abiotic and biotic stresses, adhesion to substrates and subsequent damage of colonized surfaces, the rock-inhabiting fungus Knufia petricola was chosen as gene functions in this fungus can be studied by CRISPR/Cas9-based genome editing. The putative melanogenic genes were identified in the genome of K. petricola, deleted to confirm their involvement in DHN melanogenesis and co-expressed in Saccharomyces cerevisiae for reconstruction of the synthesis pathway. Phenotypes of DHN-deficient mutants are studied. Here, we will discuss the role of the DHN melanin layer on the outer cell wall in tolerating UV irradiation. T2 - Annual conference 2023 of the Association for General and Applied Microbiology CY - Göttingen, Germany DA - 10.09.2023 KW - Pigment KW - UV radiation KW - Tolerance PY - 2023 AN - OPUS4-58425 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 JF - Fungal Biology 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 - CONF A1 - Schumacher, Julia A1 - Brandhorst, Antonia A1 - Nitsche, Sarah A1 - Erdmann, Eileen A1 - Gerrits, Ruben A1 - Gorbushina, Anna T1 - The roles of DHN melanin and the stress-activated MAP kinase in the rock inhabitant Knufia petricola N2 - Black fungi/yeasts exhibit high stress tolerance, yeast‑like or meristematic growth, and constitutive 1,8-dihydroxynaphthalene (DHN) melanin formation. Due to their slow growth, robust cell walls and the lack of sexual cycles and genetic tools, the underlying mechanisms of their phenotypic traits have remained largely unexplored. Using recently developed genetic tools, it is now possible to manipulate the genome of the rock-inhabiting model fungus Knufia petricola. Thus, gene functions and the cell biology of black fungi can be studied using CRISPR/Cas9-based genome editing and live-cell imaging with genetically encoded fluorescent proteins. Here, we are addressing the question to which extent constitutive pigment formation (melanin and carotenoids) and responses mediated by the stress-activated mitogen-activated protein (MAP) kinase contribute to the observed extremotolerance of K. petricola. The mutations of pks1, phs1 and both genes result in melanin-free (pink), carotenoid-free (black) and pigment-free (white) strains, respectively. The other putative melanogenic genes were identified in the genome, deleted to confirm their involvement in DHN melanogenesis and co-expressed in Saccharomyces cerevisiae for reconstruction of the synthesis pathway. Sak1 encoding the stress-activated MAP kinase was deleted in the wild-type and different pigment-deficient backgrounds. Growth of the obtained single, double and triple deletion mutants was tested by droplet tests on media supplemented with different stress-inducing agents. The Δsak1 mutants show slightly reduced growth rates even without environmental pressure and are hypersensitive to different stresses: e.g. osmotic, oxidative, membrane, pH and heat stress. Melanin-free Δsak1 mutants are more sensitive than black Δsak1 mutants to some but not all stress conditions, suggesting that melanin and the SAK1 pathway have complementary roles in protecting K. petricola from stress. T2 - 16th European Conference on Fungal Genetics CY - Innsbruck, Austria DA - 05.03.2023 KW - Black fungus KW - Extremotolerance KW - Pigments PY - 2023 AN - OPUS4-57147 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 JF - npj Materials degradation 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 - 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 JF - Life 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 - Schumacher, Julia ED - Scott, B. ED - Mesarich, C. T1 - Role of light in the life cycle of Botrytis cinerea T2 - Plant Relationships. The Mycota N2 - The fungus Botrytis cinerea (Botryotinia fuckeliana) infects more than 500 plant species and causes a wide range of symptoms: soft rots, accompanied by collapse and water-soaking of tissues followed by the appearance of gray masses of conidia on leaves and soft fruits (gray mold), and spots that may turn brown to full-scale soft rotting on flower petals (Botrytis blight). In general, B. cinerea is responsible for severe economic losses that are either due to the damage of growing plants in the field or the rot of harvested fruits, flowers, and vegetables during storage under cold and humid conditions. B. cinerea has adapted to the plant host and its environment by evolving strategies to use plant tissues for proliferation in terms of a necrotrophic lifestyle, and to survive biotic stresses (host responses) as well as abiotic factors of the host’s environment such as sunlight and concomitant stresses. B. cinerea maintains a complex regulatory network of light-sensitive proteins and signal transduction pathways to use light for coordinating stress responses, virulence, and reproduction. Different light-controlled reproduction cycles enable B. cinerea to live in moderate climate zones by infecting and propagating in summer and resting in winter when green host tissues are unavailable. KW - Gray mold fungus KW - Plant pathogen KW - Light KW - Photoreceptors KW - Development PY - 2023 DO - https://doi.org/10.1007/978-3-031-16503-0_14 VL - 5 SP - 329 EP - 346 PB - Springer, Cham AN - OPUS4-56724 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schumacher, Julia T1 - Rock-EATING FUNGI - Genetic Dive into the Biology of the Microcolonial 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. The 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. T2 - VAAM-Jahrestagung 2021 CY - Online meeting DA - 18.03.2021 KW - Fungus KW - Genetics KW - Marble PY - 2021 AN - OPUS4-52496 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schumacher, Julia T1 - Photoregulation of Metabolism in Plant- & Rock-associated Black Fungi N2 - Fungi that share light-flooded habitats with phototrophs may profit from excess photosynthetic products. But to cope with sunlight-associated stresses it is important for fungi to accurately sense and respond to changes in light. To test the hypothesis that light is an environmental cue that Ascomycota use to coordinate growth, stress responses as well as to establish pathogenic or symbiotic relationships, the photoreceptor (PR) distribution in species from different ecological niches was analysed. The genomes of black fungi from phyllosphere and exposed solid surfaces contain multiple photoreceptors (PRs). The filamentous foliar plant pathogen Botrytis cinerea (Leotiomycetes) has a highly sophisticated photosensory and signalling system that helps to avoid light and to locate susceptible hosts. Rock-inhabiting Dothideo- and Eurotiomycetes including Knufia petricola possess equal numbers of PRs along with the same set of protective pigments. This similarity between black fungi from plant and rock surfaces suggests that photoperception and photoregulation are important for fungi that avoid loss of energy and nutrients through cooperation with phototrophs. T2 - European Fungal Secondary Metabolism Symposium CY - Hannover, Germany DA - 30.09.2019 KW - Fungi KW - Secondary metabolism KW - Light regulation KW - Pigments PY - 2019 AN - OPUS4-49638 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schumacher, Julia T1 - Photoregulation in Plant- & Rock-associated Black Fungi N2 - Fungi that share light-flooded habitats with phototrophs may profit from excess photosynthetic products. But to cope with sunlight-associated stresses it is important for fungi to accurately sense and respond to changes in light. To test the hypothesis that light is an environmental cue that Ascomycota use to coordinate growth, stress responses as well as to establish pathogenic or symbiotic relationships, the photoreceptor (PR) distribution in species from different ecological niches was analysed. The genomes of black fungi from phyllosphere and exposed solid surfaces contain multiple photoreceptors (PRs). The filamentous foliar plant pathogen Botrytis cinerea (Leotiomycetes) has a highly sophisticated photosensory and signalling system that helps to avoid light and to locate susceptible hosts. Rock-inhabiting Dothideo- and Eurotiomycetes including Knufia petricola possess equal numbers of PRs along with the same set of protective pigments. This similarity between black fungi from plant and rock surfaces suggests that photoperception and photoregulation are important for fungi that avoid loss of energy and nutrients through cooperation with phototrophs. T2 - Scientific Colloquium at the Department of Genetics, University of Seville CY - Seville, Spain DA - 24.10.2019 KW - Light sensing KW - Knufia petricola KW - Botrytis cinerea PY - 2019 AN - OPUS4-49637 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schumacher, Julia T1 - Photoperception in plant- and rock-associated black fungi N2 - Fungi that share light-flooded habitats with phototrophs may profit from their excess photosynthetic products. Sunlight-associated stresses are however multiple: high temperatures, UV radiation with associated DNA damage, accumulation of reactive oxygen species (ROS), desiccation and osmotic stresses. Ascomycota dominating light-flooded habitats accurately sense and respond to changes in light using it as a cue to coordinate growth, stress responses as well as to establish pathogenic or symbiotic relationships. Two species from two light-flooded habitats – phyllosphere and sun-exposed solid surfaces – were analysed for their photoreceptor distribution. In both habitats phototroph-associated and black [dihydroxynaphthalene (DHN) melanin-containing] fungi are prevalent. This diversity was sampled with the plant-associated fungus Botrytis cinerea (Leotiomycetes), while Knufia petricola (Eurotiomycetes) was included as a typical biofilm-former on sun-exposed solid surfaces e.g. rocks, building facades, roofs, and solar panels. The analysis has shown that genomes of black fungi contain more photoreceptors than animal pathogens and saprophytes such as Aspergillus nidulans and Neurospora crassa1,2. B. cinerea that causes the grey mould disease by infecting the above-ground parts of more than 200 dicots has a highly sophisticated photosensory and signalling system that helps to avoid light and to locate susceptible hosts1. Rock-inhabiting Dothideomycetes and Eurotiomycetes including Knufia petricola possess equal numbers of photoreceptors along with the same set of protective metabolites i.e. melanin, carotenoids and mycosporines2. This similarity between black fungi from plant and rock surfaces suggests that photoperception and -regulation are important for sun-stressed fungi that receive nutrients through cooperation with phototrophs. CRISPR/Cas9-based genetic tools for manipulating K. petricola were established3 and are currently used for elucidating the functions of the different photoreceptors in the biology of rock-inhabiting fungi. This work was supported by the grant SCHU 2833/4-1 from the German Research Foundation (DFG) and internal funds of the BAM. T2 - 19th Congress of the European Society for Photobiology CY - Online meeting DA - 30.08.2021 KW - Fungi KW - Light KW - Extreme environments PY - 2021 AN - OPUS4-53191 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -