TY - CONF A1 - Stohl, Leonie A1 - von Werder, Julia T1 - Funktionalisierter Beton: Biorezeptivität von strukturiertem UHPC N2 - Understanding the fundamentals of bioreceptivity enables the developement of functionalized materials. Concrete as the most used building material worldwide is of special interest as microbially greened panels may represent an alternative to classic façade greening with plants. As standards are not ye established, material characterization in context as well as bioreceptivity assessment itself has been proven to be difficult. The presentation reviews first results of methods adapted for this application. T2 - Tagung des Instituts für Bioanalytik: Bewertung biozidhaltiger Baustoffe CY - Augsburg, Germany DA - 29.06.2023 KW - Biorezeptivität KW - Beton KW - UHPC KW - Biofilm PY - 2023 AN - OPUS4-58375 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Catanzaro, Ilaria A1 - Nitsche, Sarah A1 - Gorbushina, Anna A1 - Schumacher, Julia A1 - Onofri, S. T1 - Generation of Cryomyces antarcticus mutants to explore the importance of DHN melanin for survival N2 - Cryomyces antarcticus, a cryptoendolithic melanized fungus endemic to Antarctica (phylum Ascomycota, class Dothideomycetes incertae sedis), has demonstrated high capability to survive extreme environmental conditions like those found in space (e.g., ionizing radiation, vacuum, microgravity), thus fueling fundamental astrobiological questions like “the search for life beyond Earth”. Its extraordinary resilience has been attributed to the presence of thick, highly melanized cell walls, which may contain both DHN and DOPA melanins. To better understand the contribution of DHN melanin to the overall resilience of C. antarcticus, we decided to generate melanin-deficient mutants by genetic engineering. For this, the melanin-PKS (polyketide synthase)- encoding ortholog was identified in the C. antarcticus CBS 116301 genome and used to design primers for re-sequencing of the capks1 locus in the strain CCFEE 515. Based on the genetic toolkit developed for the black fungus Knufia petricola, we designed a strategy for mutating capks1. Protoplasts of C. antarcticus CCFEE 515 were generated and co-transformed with a circular AMA-containing plasmid for expression and in-vivo assembly of Cas9 and two capks1-specific sgRNAs and a PCR-generated donor DNA i.e., a hygromycin resistance cassette flanked by ~75-bp-long sequences homologous to the capks1 locus. Transformation of C. antarcticus is challenging because of its very slow growth and uncertain gene copy number. It took six months from obtaining enough biomass for cell wall lysis to transferring the putatively resistant transformants for genotyping, but we eventually managed to generate three independent non-melanized (whitish) Δcapks1 mutants! We are now awaiting first complemented strains as ultimate genetic transformation control. They are considered to have a wild-type-like pigmentation due to the reintroduction of the wild-type capks1 into the Δcapks1 mutant. T2 - IUBMB Focused Meeting on Extremophilic Fungi (FUN-EX) CY - Ljubljana, Slovenia DA - 19.09.2023 KW - Black fungus KW - Genetics KW - UV tolerance KW - Melanin PY - 2023 AN - OPUS4-58458 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schumacher, Julia A1 - Erdmann, Eileen A. A1 - Nitsche, Sarah A1 - Gerrits, Ruben A1 - Heeger, Felix A1 - Gorbushina, Anna T1 - Genetic engineering of black fungi: lessons learned from Knufia petricola N2 - The exponential rise in the number of fungal genomes sequenced by next-generation sequencing techniques makes it necessary to increase efforts to correctly annotate and assign gene functions. There are two possibilities to explore a genome and its gene functions. The hypothesis-based method proves the function of already existing gene/allele candidates by targeted mutagenesis - so called reverse genetics. The basis of forward genetics approaches is the random mutagenesis of the genome, followed by screening of obtained mutants for the phenotype of interest, and identification of the mutated genes in the respective mutants. This strategy is hypothesis-generating, means it is necessary to verify the relationship between the detected mutations and the observed phenotype by targeted mutagenesis of the identified gene. We developed a toolbox for editing the genome of the rock inhabitant Knufia petricola [Eurotiomycetes, Chaetothyriales] that allows the study of the phenotypic characteristics of black fungi such as the regulation of pigment synthesis, general stress responses, oligotrophy, and the unusual modes of cell division by advanced reverse and forward genetics approaches. The toolbox includes the annotated genome sequence of strain A95, efficient strategies for CRISPR/Cas9-based genome editing and live-cell imaging using genetically encoded fluorescent proteins, as well as protocols for -omics approaches and for simulation of mineral weathering. A forward genetics approach using transposon mutagenesis is currently developed for identifying essential genes. The established protocols and knowledge gained from K. petricola form a starting point for making other fungi from extreme environments accessible to genetic manipulation. T2 - IUBMB Focused Meeting on Extremophilic Fungi (FUN-EX) CY - Ljubljana, Slovenia DA - 19.09.2023 KW - Fungus KW - Extremotolerance KW - Genetics KW - Model organism PY - 2023 AN - OPUS4-58424 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schumacher, Julia T1 - Genetic manipulation of the microcolonial black fungus Knufia petricola N2 - Microcolonial black fungi, a polyphyletic group of ascomycetes, exhibit constitutive melanin formation, yeast-like growth and high stress tolerances. They dominate – often together with bacteria and algae in sub-aerial biofilms – a range of hostile environments including natural and man-made ones, from salterns to dishwashers, roofs and solar panels. Because of lacking genetic tools and the slow growth of most isolates, the genetic bases for these specific properties are largely unknown. The rock-inhabiting fungus Knufia petricola (Eurotiomycetes, Chaetothyriales) exhibits all characteristics of microcolonial black fungi and was selected as recipient for genetic engineering to study gene functions and genetic interactions. Different variants of green and red fluorescent proteins were successfully expressed indicating that fluorescence microscopy using genetically encoded fluorescent proteins and fluorescent dyes enables various cell biology approaches. Furthermore, genes of biosynthetic pathways (DHN melanin, carotenoids, uracil, adenine) were successfully mutated by applying traditional gene replacement and plasmid-based or ribonucleoprotein (RNP)-based CRISPR/Cas9 or silenced by RNA interference (RNAi). The availability of this advanced and efficient genetic toolbox and the annotated genome sequence of strain A95 makes K. petricola an excellent model for exploring the secrets of microcolonial black fungi. T2 - Departmental colloquium of the Tuscia University CY - Viterbo, Italy DA - 14.02.2020 KW - black fungus KW - genetics KW - Crispr/Cas9 PY - 2020 AN - OPUS4-50592 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schlegel, Moritz-Caspar T1 - Gut Ding braucht Weile - Ressourceneffizienz im Ökodesign und das „Winterpaket“ N2 - Die Kreislaufwirtschaftsinitiative der Europäischen Kommission fordert klar grundlegende Aspekte der Kreislaufwirtschaft in bereits bestehende Rechtsrahmen einzubetten. Anforderungen zur gesetzlich vorgeschriebenen Lebensdauer, Reparierbarkeit und Recyclingfähigkeit von Produkten sollen zukünftig auf europäischer Ebene ins Leben gerufen werden. Ein geeignetes Spielfeld ist die Ökodesign-Richtlinie. Momentan regelt sie primär die Energieeffizienz von energieverbrauchsrelevanten Produkten, doch der Fokus wird aktuell um Anforderungen an den nachhaltigen Einsatz von Ressourcen erweitert. Bei den gegenwärtigen Aktivitäten im Ökodesign steht der Normungsauftrag M/543 im Zentrum. Es wurde 2015 von der Europäischen Kommission an CEN/CENELEC vergeben und später stieß auch ETSI dazu. Bislang sind acht generische EN-Normen und zwei Fachberichte (Technical Report (TR)) geplant. Sie sollen grundlegende Aspekte der Nachhaltigkeit von energieverbrauchsrelevanten Produkten, die von der Ökodesign-Richtlinie betroffen sind, abdecken. Trotz der zu erwartenden hohen Reichweite dieses Vorhabens scheinen die Inhalte und der eigentliche Zweck der zu erarbeitenden Normen vielen Marktakteuren noch nicht vollständig bekannt zu sein. Daher hat ein nationaler Zentralverband eingeladen zu berichten. In dem Vortrag werden Antworten auf folgende Fragen gegeben: Warum im Ökodesign? Was ist geplant? Was ist betroffen? Worum geht es in den Normen? T2 - AK-Sitzung CY - ZVEI, Frankfurt a. Main, Germany DA - 19.09.2018 KW - Normung KW - Ressourceneffizienz KW - Ökodesign KW - Kreislaufwirtschaft PY - 2018 AN - OPUS4-45992 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Altmann, Korinna T1 - Harmonisation efforts of the CUSP projects N2 - The talk is about harmonisation approaches within CUSP. First the five H2020 projects for research on risk assessment regarding microplastics topic were presented. Secondly, harmonisation process were discussed. T2 - VAMAS SC 47 Meeting CY - Turin, Italy DA - 19.10.2022 KW - Microplastics KW - Harmonisation KW - Nanoplastics KW - H2020 KW - Risk assessment PY - 2022 AN - OPUS4-56077 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Wilke, Olaf T1 - Harmonised test standard EN 16516 - common approach for all building products for interiors N2 - The presentation gives general information about the European Standard EN 16516 “Construction products – Assessment of release of dangerous substances – Determination of emissions into indoor air”. This test standard was developed based on the mandate M/366 of the European commission and is a horizontal reference method for the determination of volatile organic compounds (VOC) from different classes of construction (building) products. Specific test conditions are to be selected by the product TCs (technical committees) in a way that a product is tested under its intended condition of use. The test is based on the use of emission test chambers which are operated at constant air change rate and climate (23°C, 50 % r.h.) over 28 days. The standard defines the conditions and requirements for the measurement including loading factor, air change rate, sampling, analysis and calculation of emission rates of the substances. A 30 m³ reference room is described which is used to calculate air concentrations from the determined emission rates. The standard EN 16516 enables the evaluation of construction products regarding their emissions into indoor air under defined and comparable conditions. The evaluation includes the determination of identified target compounds, non-identified target compounds, volatile carcinogenic compounds and the sum values TVOC, TSVOC and R. T2 - UBA Conference - Limiting health impacts of construction products regarding VOC CY - Online meeting DA - 20.04.2021 KW - Building products KW - EN 16516 KW - Emisson testing KW - Construction products PY - 2021 AN - OPUS4-58079 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Altmann, Korinna A1 - Braun, Ulrike T1 - Herausforderungen bei der Mikroplastik-Analyse: Probennahme, Probenvorbereitung und Analyse N2 - Seit einigen Jahren wird in den Medien viel über das Thema Mikroplastik (MP) diskutiert. Dabei wird häufig von einer großen Anzahl an MP-Partikeln in allen Bereichen der Umwelt, wie dem Wasser, dem Boden, der Luft und in Biota sowie über mögliche Gefahren ausgehend von MP, berichtet. Jedoch sind diese Studien häufig nicht untereinander vergleichbar, da verschiedene Konzepte der Probennahme, der Aufbereitung und unterschiedliche Detektionsverfahren verwendet werden. Um diesen Mangel an vergleichbaren und aussagekräftigen Daten zu beheben, wird aktuell versucht, eine Harmonisierung der Mikroplastikanalytik voranzutreiben. Neue, schnellere Methoden zur MP Quantifizierung wurden entwickelt. Gleichzeitig werden neue Strategien zur Probenahme und Probenvorbereitung konzipiert, da hier die Fehlerquellen besonders große Auswirkungen auf das Ergebnis haben. In diesem Vortrag werden verschiedene Verfahren bezogen auf Probennahme, -vorbereitung und Analytik gezeigt und bezüglich ihres möglichen Ergebnisses diskutiert. T2 - 7. Workshop Technologieplattform Mikroverkapselung - Bioabbaubare Mikropartikel und Mikrokapseln CY - Bremen, Germany DA - 24.10.2019 KW - Mikroplastik KW - Analytik PY - 2019 AN - OPUS4-49395 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schumacher, Julia T1 - How light determines the life of the plant pathogen Botrytis cinerea N2 - The rotation of the Earth causes a day/night cycle that is characterized by changing light and temperature conditions. Fungi actively sense the environmental light conditions to induce protective mechanisms against the light-associated stresses and to regulate development. Since fungi adapted to habitats with different light regimes, the complexities of their ‘visual’ systems and photoresponses may vary significantly. Fungi that are associated with plants experience a special light regime because the host seeks optimum light conditions for photosynthesis – and the fungus must cope with them. Moreover, fungi living under the canopy are confronted with an altered spectrum enriched for green and far-red light. Plants sense light to coordinate growth and morphogenesis: a low red:far-red ratio indicates the presence of competitors (shading by other plants) and triggers the shade avoidance response which is accompanied by increased susceptibility to nectrophic pathogens. Botrytis cinerea, the causal agent of gray mold diseases on many plant species in moderate climate zones (high humidity, temperatures around 20°C, seasons), exhibits striking photoresponses accompanied by a broad action spectrum and a high number of photoreceptors. Light is the most important factor controlling morphogenesis: it induces conidiation for disease spreading (summer cycle) and represses sclerotial development for survival and/or sexual recombination (winter cycle). In the past years, we identified several cellular components involved in photoperception and regulation of photomorphogenesis and virulence indicating that the same signaling pathways are of relevance for both processes, propagation and infection. Notably, we recognized the role of the phytochromes (red/far-red light sensors) for coordinated responses to light and elevated temperatures (photo-/thermomorphogenesis) – similar to the function of Arabidopsis PhyB. BcPHY2-like thermosensors are likely restricted to the Leotiomycetes, which include several plant pathogens of the moderate climate zones. T2 - Fungal Genetics Conference 2019 CY - Asilomar, CA, USA DA - 12.03.2019 KW - fungus KW - light KW - plant pathogen KW - signaling KW - virulence PY - 2019 AN - OPUS4-48128 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Oberbeckmann, Sonja A1 - Scales, Brittan A1 - Hassenrück, Christiane A1 - Moldaenke, Lynn A1 - Rummel, Christoph A1 - Völkner, Corinna A1 - Hassa, Julia A1 - Rückert, Christian A1 - Rynek, Robby A1 - Busche, Tobias A1 - Kalinowski, Jörn A1 - Jahnke, Annika A1 - Schmitt-Jansen, Mechthild A1 - Wendt-Potthoff, Katrin T1 - Hunting for pigments in plastic biofilms from the Great Pacific Garbage Patch N2 - Each year, an immense amount of plastic debris enters marine ecosystems, much of which ends up in the Great Pacific Garbage Patch (GPGP). The plastic fragments are home to a diverse community of microorganisms, and while researchers have gained a better understanding of these marine plastic biofilms, we lack insights into the physiology and genomic potential of the bacteria that colonize them. Examining the lifestyle of plastic colonizers from the GPGP is particularly intriguing, as the high concentration of plastics in this accumulation zone might allow for a microbial adaptation to this unique man-made habitat. A range of pigmentation was observed in 67 isolated strains obtained directly from plastic pieces sampled from the GPGP surface water. Sequence comparison between the cultivated bacterial strains and the 16S rRNA gene amplicon dataset confirmed that most of the cultivates could also be captured through DNA-only methods. Whole genome analysis of four taxonomically diverse representatives revealed multiple carotenoid pathways, including those to produce less common glycosylated carotenoids, like sarcinaxanthin glucoside. Further, we identified a potentially new Rhodobacteraceae species containing a photosynthetic gene cluster (PGC). Absorption analysis confirmed the actual production of the carotenoids and bacteriochlorophyll a. Floating plastics represent a habitat with strong UV-light exposure, making the protection with antioxidant carotenoids as well as the ability to use light as an energy source highly beneficial traits for plastic colonizers. Our findings indicate, that the production of pigments is a common adaption mechanism for plastic-associated bacteria, and that plastic biofilms present a so far overlooked source of rare carotenoids and light-harvesting mechanisms. T2 - Annual Conference of The Association for General and Applied Microbiology (VAAM) CY - Göttingen, Germany DA - 10.09.2023 KW - Plastic biofilm KW - Phototrophic bacteria KW - Plastic pollution PY - 2023 AN - OPUS4-58443 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -