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Erscheinungsjahr
- 2020 (1334) (entfernen)
Dokumenttyp
- Zeitschriftenartikel (622)
- Vortrag (398)
- Beitrag zu einem Tagungsband (99)
- Posterpräsentation (81)
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Schlagworte
- Nanoparticles (45)
- Korrosion (29)
- Additive manufacturing (27)
- Fluorescence (27)
- Corrosion (25)
- Nanoparticle (18)
- Laser beam welding (15)
- Additive Manufacturing (14)
- Electron microscopy (14)
- Laser-induced periodic surface structures (LIPSS) (14)
Organisationseinheit der BAM
- 6 Materialchemie (284)
- 1 Analytische Chemie; Referenzmaterialien (219)
- 8 Zerstörungsfreie Prüfung (213)
- 7 Bauwerkssicherheit (195)
- 9 Komponentensicherheit (151)
- 4 Material und Umwelt (135)
- 5 Werkstofftechnik (110)
- 3 Gefahrgutumschließungen; Energiespeicher (93)
- 6.3 Strukturanalytik (84)
- 6.1 Oberflächen- und Dünnschichtanalyse (81)
Paper des Monats
- ja (17)
Thermographic reconstruction of defects that lie in the bulk of a sample is a difficult task because entropy production during heat diffusion leads to information loss. To reconstruct defects one has to solve an inverse heat conduction problem. The quality of the reconstruction is closely related to the information content of the observed data set that is reflected by the decreasing ability to spatially resolve a defect with growing defect depth. In this work we show a 2D reconstruction of rectangular slots with different width-to-depth ratios in a metallic sample. For this purpose, we apply the virtual wave concept and incorporate positivity and sparsity as prior information to overcome the diffusion-based information loss partially. The reconstruction is based on simulated and experimental pulse thermography data. In the first reconstruction step, we compute a virtual wave field from the surface temperature data. This allows us, in the second step, to use ultrasonic backpropagation methods for image reconstruction.
Die chemische Analyse von Beton gehört bei der Ermittlung des Ist-Zustandes von Stahl- und Spannbetonbauwerken zu den grundlegenden Untersuchungen bei der Zustandserfassung. Der tiefenabhängige Chloridgehalt sowie der Karbonatisierungsfortschritt müssen bei der Bewertung des Ist-Zustandes, beim Instandsetzungsbedarf und ggf. bei der Planung eines erforderlichen Instandsetzungskonzeptes mit einbezogen werden. Seit Mitte der 1990er Jahre wird an der Bundesanstalt für Materialforschung und -prüfung die laserinduzierte Plasmaspektroskopie (engl. Laser-induced Breakdown Spectroscopy) – kurz LIBS – stetig für die chemische Analyse von Beton weiterentwickelt. In diesem Beitrag wird der aktuelle Stand der Forschung/ Technik und der Normungsarbeit präsentiert, sowie zukünftig mögliche Anwendungen vorgestellt. Mit den aktuellen LIBS-Systemen (mobiles LIBS-System sowie kommerziell erhältliche Laborsysteme) ist es möglich den Chloridgehalt ortsaufgelöst in wenigen Minuten zu quantifizieren, sogenannte Elementlandkarten zu erstellen und somit die Elementverteilung innerhalb des Betons zu visualisieren. Durch das Scannen eines Bohrkernquerschnittes und der simultanen Erfassung von mehreren Elementen mit einer Messung, ist es zudem möglich, die Gesteinskörnung in den Messdaten zu identifizieren und den Chloridgehalt auf die Zementsteinmatrix zu beziehen. Fehlerquellen aus der Umrechnung der betonbezogenen Chloridgehalte können dadurch minimiert werden. Neben den Grundlagen von LIBS werden vor allem aktuelle Praxisbeispiele vorgestellt, die die aktuellen Möglichkeiten des Verfahrens veranschaulichen.
Heterodyne Eddy Current Testing Using Magnetoresistive Sensors for Additive Manufacturing Purposes
(2020)
In recent years additive manufacturing technologies have become widely popular. For complex functional components or low volume production of workpieces, laser powder bed fusion can be used. High safety requirements, e.g. in the aerospace sector, demand extensive quality control. Therefore, offline non-destructive testing methods like computed tomography are used after manufacturing. Recently, for enhanced profitability and practicality online non-destructive testing methods, like optical tomography have been developed. This paper discusses the applicability of eddy current testing with magnetoresistive sensors for laser powder bed fusion parts. For this purpose, high spatial resolution giant magnetoresistance arrays are utilized for testing in combination with a single wire excitation coil. A heterodyne principle minimizes metrology efforts. This principle is compared to conventional signal processing in an eddy current testing setup using an aluminum test sample with artificial surface defects. To evaluate the influence of the powder used in the manufacturing process on eddy current testing and vice versa, a laser powder bed fusion mock-up made from stainless steel powder (316L) is used with artificial surface defects down to 100 µm. This laser powder bed fusion specimen was then examined using eddy current testing and the underlying principles.
The erosion of natural sediments by a superficial fluid flow is a generic situation in many usual geological or industrial contexts. However, there is still a lack of fundamental knowledge about erosional processes, especially concerning the role of internal cohesion and adhesive stresses on issues such as the critical flow conditions for the erosion onset or the kinetics of soil mass loss. This contribution investigates the influence of cohesion on the surface erosion by an impinging jet flow based on laboratory tests with artificially bonded granular materials. The model samples are made of spherical glass beads bonded either by solid bridges made of resin or by liquid bridges made of a highly viscous oil. To quantify the intergranular cohesion, the capillary forces of the liquid bridges are here estimated by measuring their main geometrical parameters with image-processing techniques and using well-known analytical expressions. For the solid bonds, the adhesive strength of the materials is estimated by direct measurement of the yield tensile forces and stresses at the particle and sample scales, respectively, with specific traction tests developed for this purpose. The proper erosion tests are then carried out in an optically adapted device that permits a direct visualization of the scouring process at the jet apex by means of the refractive index matching technique. On this basis, the article examines qualitatively the kinetics of the scour crater excavation for both scenarios, namely, for an intergranular cohesion induced by either liquid or solid bonds. From a quantitative perspective, the critical condition for the erosion onset is discussed specifically for the case of the solid bond cohesion. In this respect, we propose here a generalized form of the Shields criterion based on a common definition of a cohesion number from yield tensile values, derived at both micro- and macroscales. The article finally shows that the proposed form manages to reconcile the experimental data for cohesive and cohesionless materials, the latter in the form of the so-called Shields curve along with some previous results of the authors which have been appropriately revisited.
Alfeld, M. ; Eckhardt, H.-S. ; Kraft, J. ; Maiwald, Michael ; Meermann, Björn ; Merz, K. ; Prikler, S. ; Richert, J. ; Steiner, G. ; von Tümpling, W.
Zusätzlich zu Methodenentwicklung, Miniaturisierung und Kopplungsverfahren zeigen sich die Hyperspektroskopie zusammen mit Imaging‐Verfahren, der Einzelmolekülnachweis und der Einsatz von 3‐D‐Druckern als neue Schwerpunkte. Hinzu kommen künstliche Intelligenz bei Sensoren, Bildgebungsverfahren und Prozesssteuerung sowie die Vernetzung von Analyse‐ und Laborgeräten. Trends und Forschungsthemen aus der analytischen Chemie, zusammengestellt von elf Autoren, koordiniert von Günter Gauglitz.
Currently, sulfate-reducing bacteria (SRB) is regarded as the main culprit of microbiologically influenced corrosion (MIC), mainly due to the low reported corrosion rates of other microorganisms. For example, the highest reported corrosion rate for methanogens is 0.065 mm/yr. However, by investigating methanogen-induced microbiologically influenced corrosion (Mi-MIC) using an in-house developed versatile multiport flow test column, extremely high corrosion rates were observed. We analyzed a large set of carbon steel beads, which were sectionally embedded into the test columns as substrates for iron-utilizing methanogen Methanobacterium IM1. After 14 days of operation using glass beads as fillers for section separation, the highest average corrosion rate of Methanobacterium IM1 was 0.2 mm/yr, which doubled that of Desulfovibrio ferrophilus IS5 and Desulfovibrio alaskensis 16109 investigated at the same conditions. At the most corroded region, nearly 80% of the beads lost 1% of their initial weight (fast-corrosion), resulting in an average corrosion rate of 0.2 mm/yr for Methanobacterium IM1-treated columns. When sand was used as filler material to mimic sediment conditions, average corrosion rates for Methanobacterium IM1 increased to 0.3 mm/yr (maximum 0.52 mm/yr) with over 83% of the beads having corrosion rates above 0.3 mm/yr. Scanning electron images of metal coupons extracted from the column showed methanogenic cells were clustered close to the metal surface. Methanobacterium IM1 is a hydrogenotrophic methanogen with higher affinity to metal than H2. Unlike SRB, Methanobacterium IM1 is not restricted to the availability of sulfate concentration in the environment. Thus, the use of the multiport flow column provided a new insight on the corrosion potential of methanogens, particularly in dynamic conditions, that offers new opportunities for monitoring and development of mitigation strategies. Overall, this study shows under certain conditions methanogenic archaea can cause higher corrosion than SRB, specific quantifications, i.e., maximum, average, and minimum corrosion rates can be determined, and that spatial statistical evaluations of MIC can be carried out.
On the influence of crystallography on creep of circular notched single crystal superalloy specimens
(2020)
The present work contributes to a better understanding of the effect of stress multiaxiality on the creep behavior of single crystal Ni-base superalloys. For this purpose we studied the creep deformation and rupture behavior of double notched miniature creep tensile specimens loaded in three crystallographic directions [100], [110] and [111] (creep conditions: 950 °C and 400 MPa net section stress). Crystal plasticity finite element method (CPFEM) was used to analyze the creep stress and strain distributions during creep. Double notched specimens have the advantage that when one notch fails, the other is still intact and allows to study a material state which is close to rupture. No notch root cracking was observed, while microstructural damage (pores and micro cracks) were frequently observed in the center of the notch root region. This is in agreement with the FEM results (high axial stress and high hydrostatic stress in the center of the notched specimen). Twinning was observed in the notch regions of [110] and [111] specimens, and <112> {111} twins were detected and analyzed using orientation imaging scanning electron microscopy. The present work shows that high lattice rotations can be detected in SXs after creep fracture, but they are associated with the high strains accumulated in the final rupture event.
Thomas, Maximilian ; Vollert, F. ; Weidemann, Jens ; Gibmeier, J. ; Kromm, Arne ; Kannengießer, Thomas
The subject of this study is how, and to what extent, Varestraint/Transvarestraint test results are influenced by both testing parameters and characteristics of evaluation methods. Several different high-alloyed martensitic LTT (low Transformation temperature) filler materials, CrNi and CrMn type, were selected for examination due to their rather distinctive solidification cracking behaviour, which aroused interest after previous studies. First, the effects of different process parameter sets on the solidification cracking response were measured using standard approaches. Subsequently, microfocus X-ray computer tomography (μCT) scans were performed on the specimens. The results consistently show sub-surface cracking to significant yet varying extents. Different primary solidification types were found using wavelength dispersive X-ray (WDX) analysis conducted on filler metals with varying Cr/Ni equivalent ratios. This aspect is regarded as the main difference between the CrNiand CrMn-type materials in matters of cracking characteristics. Results show that when it comes to testing of modern highperformance alloys, one set of standard Varestraint testing parameters might not be equally suitable for all materials. Also, to properly accommodate different solidification types, sub-surface cracking has to be taken into account.
The interaction of different charged polymers, namely anionic polycarboxylate
superplasticizer (PCE) and neutral polyethylene glycol (PEG) with potassium ions, and their effect on the yield stress of highly concentrated glass bead suspension (GBS), were studied under different concentrations of potassium ions ([K+]). It was found that, compared to the neutral PEG, the negatively charged PCE can be adsorbed on glass beads (GB), and then decreases the yield stress of GBS. The increasing concentration of free polymer in the interstitial liquid phase with the increased polymer dosage leads to the higher yield stress of GBS, which may be caused by the higher Depletion force. In addition, this effect is also related to the charge density of the polymer and the [K+] in the solution. Along with the increase in [K+], the yield stress of GBS increases significantly with the addition of PCE, but this cannot be observed with PEG, which indicates that potassium ions can interact with negatively charged PCE instead of the neutral PEG. At last, the interparticle Forces between two single GB with adsorbed PCE in solutions containing [K+] and PCE were measured by colloidal probe atomic force microscopy to better understand the interaction of the charged polymer with counterions.
Khrapov, D ; Koptyug, A. ; Manabaev, K. ; Léonard, Fabien ; Mishurova, Tatiana ; Bruno, Giovanni ; Cheneler, D. ; Loza, K. ; Epple, M. ; Surmenev, R. ; Surmeneva, M.
An ultrasonic vibration post-treatment procedure was suggested for additively manufac-tured lattices. The aim of the present research was to investigate mechanical properties andthe differences in mechanical behavior and fracture modes of Ti6Al4V scaffolds treated withtraditional powder recovery system (PRS) and ultrasound vibration (USV). Scanning electronmicroscopy (SEM) was used to investigate the strut surface and the fracture surface mor-phology. X-ray computed tomography (CT) was employed to evaluate the inner structure,strut dimensions, pore size, as well as the surface morphology of additively manufacturedporous scaffolds. Uniaxial compression tests were conducted to obtain elastic modulus,compressive ultimate strength and yield stress. Finite element analysis was performedfor a body-centered cubic (BCC) element-based model and for CT-based reconstructiondata, as well as for a two-zone scaffold model to evaluate stress distribution during elasticdeformation. The scaffold with PRS post treatment displayed ductile behavior, while USVtreated scaffold displayed fragile behavior. Double barrel formation of PRS treated scaffoldwas observed during deformation. Finite element analysis for the CT-based reconstructionrevealed the strong impact of surface morphology on the stress distribution in comparisonwith BCC cell model because of partially molten metal particles on the surface of struts,which usually remain unstressed.
Afantitis, A. ; Melagraki, G. ; Isigonis, P. ; Tsoumanis, A. ; Varsou, D. D. ; Valsami-Jones, E. ; Papadiamantis, A. ; Ellis, L.-J. A. ; Sarimveis, H. ; Doganis, P. ; Karatzas, P. ; Tsiros, P. ; Liampa, I. ; Lobaskin, V. ; Greco, D. ; Serra, A. ; Kinaret, P. A. S. ; Saarimäki, L. A. ; Grafström, R. ; Kohonen, P. ; Nymark, P. ; Willighagen, E. ; Puzyn, T. ; Rybinska-Fryca, A. ; Lyubartsev, A. ; Jensen, K. A. ; Brandenburg, J. G. ; Lofts, S. ; Svendsen, C. ; Harrison, S. ; Maier, D. ; Tamm, K. ; Jänes, J. ; Sikk, L. ; Dusinska, M. ; Longhin, E. ; Rundén-Pran, E. ; Mariussen, E. ; El Yamani, N. ; Unger, Wolfgang ; Radnik, Jörg ; Tropsha, A. ; Cohen, Y. ; Lesczynski, J. ; Hendren, C. O. ; Wiesner, M. ; Winkler, D. ; Suzuki, N. ; Yoon, T. H. ; Choi, J.-S. ; Sanabria, N. ; Gulumian, M. ; Lynch, I.
Nanotechnology has enabled the discovery of a multitude of novel materials exhibiting unique physicochemical (PChem) properties compared to their bulk analogues. These properties have led to a rapidly increasing range of commercial applications; this, however, may come at a cost, if an association to long-term health and environmental risks is discovered or even just perceived. Many nanomaterials (NMs) have not yet had their potential adverse biological effects fully assessed, due to costs and time constraints associated with the experimental assessment, frequently involving animals. Here, the available NM libraries are analyzed for their suitability for integration with novel nanoinformatics approaches and for the development of NM specific Integrated Approaches to Testing and Assessment (IATA) for human and environmental risk assessment, all within the NanoSolveIT cloud-platform. These established and well-characterized NM libraries (e.g. NanoMILE, NanoSolutions, NANoREG, NanoFASE, caLIBRAte, NanoTEST and the Nanomaterial Registry (>2000 NMs)) contain physicochemical characterization data as well as data for several relevant biological endpoints, assessed in part using harmonized Organisation for Economic Co-operation and Development (OECD) methods and test guidelines.
Integration of such extensive NM information sources with the latest nanoinformatics methods will allow NanoSolveIT to model the relationships between NM structure (morphology), properties and their adverse effects and to predict the effects of other NMs for which less data is available. The project specifically addresses the needs of regulatory agencies and industry to effectively and rapidly evaluate the exposure, NM hazard and risk from nanomaterials and nano-enabled products, enabling implementation of computational ‘safe-by-design’ approaches to facilitate NM commercialization.
Laser material processing is an emerging technology that generates surface functionalities on the basis of optical, mechanical, or chemical properties. In the form of laser surface texturing (LST), it has attracted a remarkable amount of research to tailor surface properties towards various tribological applications. The main advantages of this single-step, laser-based technology are the contactless machining, featuring a high flexibility, efficiency, and speed, along with the excellent quality of the processed products. LST can be applied precisely localized to sub-micrometric areas, but, via laser beam scanning, it is also feasible for structuring large surface areas at the square meter size.
This Special Issue focuses on the latest developments concerning the tribological performance of laser-generated periodic surface nano- and microstructures and their applications. This includes the laser-based processing of different surface patterns, such as “self-organized” laser-induced periodic surface structures (LIPSS, ripples), grooves, micro-spikes, hierarchical hybrid nano-/micro-structures, microfeatures generated by direct laser interference patterning (DLIP), or even dimples or other topographic geometries shaped by direct laser modification or ablation. The applications of these periodically nano- and micro-patterned surfaces may improve the lubricated or non-lubricated tribological performance of surfaces in conformal and even non-conformal contact through a reduction of wear, a variation of the coefficient of friction, altered load carrying capacity, etc., resulting in energy saving, improved reliability, increased lifetimes as well as durability, leading in turn to extended maintenance intervals/reduced down-time. This can be beneficial in terms of bearings, gears, engines, seals, cutting tools, or other tribological components. Fundamental aspects addressed may involve the investigation of the relevant physical and chemical effects accompanying the laser-generated nano- and microscale topographies, such as alterations of the material structures, the hardness, superficial oxidation, the role of additives contained in lubricants, surface wettability, micro-hydrodynamic effects, etc.
For this Special Issue we aim to attract both academic and industrial researchers and would like to provide a bridge between research in the fields of tribology and laser material processing in order to foster the current knowledge and present new ideas for future applications and new technologies.
Laser material processing is an emerging technology that generates surface functionalities on the basis of optical, mechanical, or chemical properties. In the form of laser surface texturing (LST), it has attracted a remarkable amount of research to tailor surface properties towards various tribological applications. The main advantages of this single-step, laser-based technology are the contactless machining, featuring a high flexibility, efficiency, and speed, along with the excellent quality of the processed products. LST can be applied precisely localized to sub-micrometric areas, but, via laser beam scanning, it is also feasible for structuring large surface areas at the square meter size.
This Special Issue focuses on the latest developments concerning the tribological performance of laser-generated periodic surface nano- and microstructures and their applications. This includes the laser-based processing of different surface patterns, such as “self-organized” laser-induced periodic surface structures (LIPSS, ripples), grooves, micro-spikes, hierarchical hybrid nano-/micro-structures, microfeatures generated by direct laser interference patterning (DLIP), or even dimples or other topographic geometries shaped by direct laser modification or ablation. The applications of these periodically nano- and micro-patterned surfaces may improve the lubricated or non-lubricated tribological performance of surfaces in conformal and even non-conformal contact through a reduction of wear, a variation of the coefficient of friction, altered load carrying capacity, etc., resulting in energy saving, improved reliability, increased lifetimes as well as durability, leading in turn to extended maintenance intervals/reduced down-time. This can be beneficial in terms of bearings, gears, engines, seals, cutting tools, or other tribological components. Fundamental aspects addressed may involve the investigation of the relevant physical and chemical effects accompanying the laser-generated nano- and microscale topographies, such as alterations of the material structures, the hardness, superficial oxidation, the role of additives contained in lubricants, surface wettability, micro-hydrodynamic effects, etc.
For this Special Issue we aim to attract both academic and industrial researchers and would like to provide a bridge between research in the fields of tribology and laser material processing in order to foster the current knowledge and present new ideas for future applications and new technologies.
Das Neutronensondenverfahren wird bereits seit vielen Jahren erfolgreich zur Eingrenzung und Quantifizierung auftretender Feuchteschäden an Fußböden eingesetzt. Hierzu bedarf es jedoch einer Vielzahl zerstörender Sondierungsbohrungen, welche die gewonnenen Messdaten kalibrieren und eine Tiefenzuordnung des Flüssigwassers zulassen. Dadurch entsteht ein zeitlicher und finanzieller Aufwand, der durch den parallelen Einsatz des elektromagnetischen Radarverfahrens vermieden werden könnte. Mit seiner hohen Sensitivität für Wasser bietet diese Messmethode die Möglichkeit der vertikalen Lokalisierung von Feuchte, was zu einer automatisierten Klassifizierung typischer Schadensfälle beitragen soll.
In einem laufenden Forschungsvorhaben werden in systematischen Laborstudien gängige Schadensfälle an häufig anzutreffenden Fußbodenaufbauten simuliert und deren Einfluss auf die genannten Verfahren untersucht. Hierbei kommen Zement- und Anhydritestriche, sowie unterschiedliche Dämmmaterialen mit variierenden Schichtdicken zum Einsatz. Wesentlicher Bestandteil der Auswertung ist die Extraktion signifikanter Signalmerkmale des Radarverfahrens, welche Rückschlüsse auf den Schadensfall und ggf. die Wassermenge zulassen. Weiterführend sollen die Kombinationsmöglichkeiten der verschiedenen Signalmerkmale und der Neutronensondendaten durch Methoden der multivariaten Datenauswertung und des maschinellen Lernens geprüft werden. Die Unabhängigkeit gegenüber wechselnden Schichtdicken und Materialien steht hierbei besonders im Fokus und soll anhand der erzielten Ergebnisse evaluiert werden.
The influence of relative humidity on friction and wear is subject of several studies in the last decades. A comprehensive understanding of physical and chemical phenomena affecting the tribology is hampered by the lack of reproducible experimental results, by the large number of variables, and by several difficulties in the detection of tribochemical processes and products.
In the present work, we analyze the wear coefficient and the wear volumes of 686 unlubricated tests performed on different oscillating tribometers with 100Cr6 balls on 100Cr6 planes at different relative humidity.
Aim of this work is to assess the repeatability and reproducibility of data, to determine the dependence of the wear coefficient on the relative humidity, to understand the underlying physicochemical phenomena and to build three dimensional maps of the wear coefficient as a function of both humidity and the product of normal force and sliding distance.
Nanoparticle suspensions were microprinted onto TEM grids for subsequent analysis by SEM/TSEM and evaluation of particle numbers using Image J software. Various nanoparticle types, concentrations and printing conditions (temperature, rel. humidity) were evaluated in order to determine the optimal conditions for producing a uniform distribution of particles on the substrate and eliminating the coffee ring effect.
Dihydroxynaphthalene (DHN) melanin is produced by different 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, survival and/or infection structures. Remarkable is the regulation of the DHN melanogenesis in the foliar plant pathogen Botrytis cinerea: it involves two differently expressed PKSs providing the precursor in conidia and sclerotia, respectively (Schumacher 2016, Mol Microbiol). In contrast, a polyphyletic group of Ascomycetes (microcolonial fungi/ black yeast) 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. As part of our continuing research on microcolonial rock-inhabiting fungi, we chose the genetically amenable Knufia petricola strain A95 (Nai et al. 2013, Fungal Genet Biol; Noack-Schönmann et al. 2014, AMB Express) for detailed studies. DHN-deficient mutants generated by targeted mutation of biosynthetic genes were studied with regard to the architecture of the cell wall and the EPS (extracellular polymeric substances) matrix, attachment to and weathering of olivine, as well as the tolerance to abiotic and biotic stresses. We will discuss the critical role of the outer cell surface (DHN melanin and EPS) in adhesion to the substrate and subsequent damage of the colonized surface.
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.
A genetic toolbox for exploring the life style of the rock-inhabiting black fungus Knufia petricola
(2020)
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.