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 U6 - 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 - Giongo, A. A1 - Favet, J. A1 - Lapanje, A. A1 - Gano, K.A. A1 - Kennedy, S. A1 - Davis-Richardson, A.G. A1 - Brown, C. A1 - Beck, A. A1 - Farmerie, W.G. A1 - Cattaneo, A. A1 - Crabb, D.B. A1 - Aung, Y.-Y. A1 - Kort, R. A1 - Brumsack, H.-J. A1 - Schnetger, B. A1 - Broughton, William J. A1 - Gorbushina, Anna A1 - Triplett, E.W. T1 - Microbial hitchhikers on intercontinental dust: high-throughput sequencing to catalogue microbes in small sand samples N2 - Microbiological studies on the intercontinental transport of dust are confounded by the difficulty of obtaining sufficient material for analysis. Axenic samples of dust collected at high altitudes or historic specimens in museums are often so small and precious that the material can only be sacrificed when positive results are assured. With this in mind, we evaluated current methods and developed new ones in an attempt to catalogue all microbes present in small dust or sand samples. The methods used included classical microbiological approaches in which sand extracts were plated out on a variety of different media, polymerase chain reaction (PCR)-based amplification of 16S/18S rRNA sequences followed by construction of clone libraries, PCR amplification of 16S rRNA sequences followed by high-throughput sequencing (HtS) of the products and direct HtS of DNA extracted from the sand. A representative sand sample collected at Bahaï Wadi in the desert of the Republic of Chad was used. HtS with or without amplification showed the most promise and can be performed on ≤100 ng DNA. Since living microbes are often required, current best practices would involve geochemical and microscopic characterisation of the sample, followed by DNA isolation and direct HtS. Once the microbial content of the sample has been deciphered, growth conditions (including media) can be tailored to isolate the micro-organisms of interest. KW - Chad KW - Deserts KW - Eukaryota KW - 16S amplicons KW - Metagenomics PY - 2013 U6 - https://doi.org/10.1007/s10453-012-9264-0 SN - 0393-5965 SN - 1573-3025 VL - 29 IS - 1 SP - 71 EP - 84 PB - Springer Science + Business Media B.V. CY - Dordrecht [u.a.] AN - OPUS4-27687 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Favet, J. A1 - Lapanje, A. A1 - Giongo, A. A1 - Kennedy, S. A1 - Aung, Y.-Y. A1 - Cataneo, A. A1 - Kort, R. A1 - Brumsack, H.-J. A1 - Schnetger, B. A1 - Triplett, E.W. A1 - Beck, A. A1 - Broughton, William J. A1 - Gorbushina, Anna T1 - Microbial hitchhikers on intercontinental dust - Chadian origins T2 - VAAM-Jahrestagung 2011 CY - Karlsruhe, Germany DA - 03.04.2011 KW - Intercontinental dust KW - Desert soil PY - 2011 SN - 0947-0867 IS - EMP114 SP - 118 PB - Spektrum, Akad. Verl. AN - OPUS4-23629 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Noack-Schönmann, Steffi A1 - Bus, T. A1 - Banasiak, Ronald A1 - Knabe, Nicole A1 - Broughton, William J. A1 - Dulk-Ras, H.D. A1 - Hooykaas, P.J.J. A1 - Gorbushina, Anna T1 - Genetic transformation of Knufia petricola A95 - a model organism for biofilm-material interactions N2 - We established a protoplast-based system to transfer DNA to Knufia petricola strain A95, a melanised rock-inhabiting microcolonial fungus that is also a component of a model sub-aerial biofilm (SAB) system. To test whether the desiccation resistant, highly melanised cell walls would hinder protoplast formation, we treated a melanin-minus mutant of A95 as well as the type-strain with a variety of cell-degrading enzymes. Of the different enzymes tested, lysing enzymes from Trichoderma harzianum were most effective in producing protoplasts. This mixture was equally effective on the melanin-minus mutant and the type-strain. Protoplasts produced using lysing enzymes were mixed with polyethyleneglycol (PEG) and plasmid pCB1004 which contains the hygromycin B (HmB) phosphotransferase (hph) gene under the control of the Aspergillus nidulans trpC. Integration and expression of hph into the A95 genome conferred hygromycin resistance upon the transformants. Two weeks after plating out on selective agar containing HmB, the protoplasts developed cell-walls and formed colonies. Transformation frequencies were in the range 36 to 87 transformants per 10 µg of vector DNA and 106 protoplasts. Stability of transformation was confirmed by sub-culturing the putative transformants on selective agar containing HmB as well as by PCR-detection of the hph gene in the colonies. The hph gene was stably integrated as shown by five subsequent passages with and without selection pressure. KW - DNA transfer KW - Fungal cell-walls KW - Protoplasts KW - Hygromycin resistance KW - Black yeast KW - Sub-aerial biofilms KW - Stress-protective morphology KW - Ancestor of opportunistic pathogens & lichens PY - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-320840 UR - http://www.amb-express.com/content/4/1/80 SN - 2191-0855 VL - 4 SP - 80, 1 EP - 6 PB - Springer CY - Heidelberg AN - OPUS4-32084 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Breitenbach, Romy A1 - Silbernagl, Dorothee A1 - Toepel, J. A1 - Sturm, Heinz A1 - Broughton, William J. A1 - Sassaki, G. L. A1 - Gorbushina, Anna T1 - Corrosive extracellular polysaccharides of the rock-inhabiting model fungus Knufia petricola N2 - Melanised cell walls and extracellular polymeric matrices protect rock-inhabiting microcolonial fungi from hostile environmental conditions. How extracellular polymeric substances (EPS) perform this protective role was investigated by following development of the model microcolonial black fungus Knufia petricola A95 grown as a sub-aerial biofilm. Extracellular substances were extracted with NaOH/formaldehyde and the structures of two excreted polymers studied by methylation as well as NMR analyses. The main polysaccharide (~ 80%) was pullulan, also known as α-1,4-; α-1,6-glucan, with different degrees of polymerisation. Αlpha-(1,4)-linked-Glcp and α-(1,6)-linked-Glcp were present in the molar ratios of 2:1. A branched galactofuromannan with an α-(1,2)-linked Manp main chain and a β-(1,6)-linked Galf side chain formed a minor fraction (~ 20%). To further understand the roles of EPS in the weathering of minerals and rocks, viscosity along with corrosive properties were studied using atomic force microscopy (AFM). The kinetic viscosity of extracellular K. petricola A95 polysaccharides (≈ 0.97 × 10-6 m2 s-1) ranged from the equivalent of 2% (w/v) to 5% glycerine, and could thus profoundly affect diffusion-dominated processes. The corrosive nature of rock-inhabiting fungal EPS was also demonstrated by its effects on the aluminium coating of the AFM cantilever and the silicon layer below. KW - Corrosion KW - EPS KW - Melanised microcolonial fungi (MCF) KW - Pullulan KW - Sub-aerial biofilms (SAB) KW - α-1,4- and α-1,6-glucans KW - AFM cantilever vibration KW - Nanoviscosity KW - Nanocorrosion of aluminium and silicon PY - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-435910 SN - 1433-4909 SN - 1431-0651 VL - 22 IS - 2 SP - 165 EP - 175 PB - Springer CY - Berlin AN - OPUS4-43591 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Nai, Corrado A1 - Wong, Helen A1 - Pannenbecker, A. A1 - Broughton, William J. A1 - Benoit, I. A1 - De Vries, R.P. A1 - Gueidan, C. A1 - Gorbushina, Anna T1 - Nutritional physiology of a rock-inhabiting, model microcolonial fungus from an ancestral lineage of the chaetothyriales (ascomycetes) N2 - Rock-inhabiting black fungi [also microcolonial or meristematic fungi (MCF)] are a phylogenetically diverse group of melanised ascomycetes with distinctive morphological features that confer extensive stress tolerance and permit survival in hostile environments. The MCF strain A95 Knufia petricola (syn. Sarcinomyces petricola) belongs to an ancestral lineage of the order Chaetothyriales (class Eurotiomycetes). K. petricola strain A95 is a rock-inhabiting MCF and its growth requirements were studied using the 96-well plate-based Biolog™ System under ~1070 different conditions (osmotic stress, pH growth optima, growth factor requirements and nutrient catabolism). A95 is an osmotolerant, oligotrophic MCF that grows best around pH 5. Remarkably, A95 shows metabolic activity in the absence of added nitrogen, phosphorus or sulphur. Correlations could be drawn between the known nutrient requirements of A95 and what probably is available in sub-aerial systems (rock and other material surfaces). Detailed knowledge of A95's metabolic requirements allowed formulation of a synthetic medium that supports strong fungal growth. KW - Knufia petricola (syn. Sarcinomyces petricola) KW - A95 KW - Chaetothyriales (Eurotiomycetes) KW - Microcolonial fungi KW - Melanised rock-inhabiting fungi KW - Biolog™ system KW - Physiological characterisation PY - 2013 U6 - https://doi.org/10.1016/j.fgb.2013.04.001 SN - 1087-1845 SN - 0147-5975 SN - 1096-0937 VL - 56 SP - 54 EP - 66 PB - Elsevier CY - Amsterdam [u.a.] AN - OPUS4-28865 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -