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 - JOUR A1 - Gay-Fraret, J. A1 - Ardissone, S. A1 - Kambara, K. A1 - Broughton, William J. A1 - Deakin, W.J. A1 - Le Quéré, A. T1 - Cyclic-Beta-glucans of Rhizobium (Sinorhizobium) sp. strain NGR234 are required for hypo-osmotic adaption, motility, and efficient symbiosis with host plants N2 - Cyclic-β-glucans (CβG) consist of cyclic homo-polymers of glucose that are present in the periplasmic space of many Gram-negative bacteria. A number of studies have demonstrated their importance for bacterial infection of plant and animal cells. In this study, a mutant of Rhizobium (Sinorhizobium) sp. strain NGR234 (NGR234) was generated in the cyclic glucan synthase (ndvB)-encoding gene. The great majority of CβG produced by wild-type NGR234 are negatively charged and substituted. The ndvB mutation abolished CβG biosynthesis. We found that, in NGR234, a functional ndvB gene is essential for hypo-osmotic adaptation and swimming, attachment to the roots, and efficient infection of Vigna unguiculata and Leucaena leucocephala. KW - Cyclic Beta-(1,2)-glucans KW - Legumes KW - Osmotic adaption KW - Swimming KW - Nodulation KW - ndvB PY - 2012 U6 - https://doi.org/10.1111/j.1574-6968.2012.02595.x SN - 0378-1097 SN - 1574-6968 VL - 333 IS - 1 SP - 28 EP - 36 PB - Wiley-Blackwell CY - Malden, Mass., USA AN - OPUS4-26111 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 - Ardissone, S. A1 - Kobayashi, H. A1 - Kambara, K. A1 - Rummel, C. A1 - Noel, K.D. A1 - Walker, G.C. A1 - Broughton, William J. A1 - Deakin, W.J. T1 - Role of BacA in lipopolysaccharide synthesis, peptide transport, and nodulation by rhizobium sp. strain NGR234 N2 - BacA of Sinorhizobium meliloti plays an essential role in the establishment of nitrogen-fixing symbioses with Medicago plants, where it is involved in peptide import and in the addition of very-long-chain fatty acids (VLCFA) to lipid A of lipopolysaccharide (LPS). We investigated the role of BacA in Rhizobium species strain NGR234 by mutating the bacA gene. In the NGR234 bacA mutant, peptide import was impaired, but no effect on VLCFA addition was observed. More importantly, the symbiotic ability of the mutant was comparable to that of the wild type for a variety of legume species. Concurrently, an acpXL mutant of NGR234 was created and assayed. In rhizobia, AcpXL is a dedicated acyl carrier protein necessary for the addition of VLCFA to lipid A. LPS extracted from the NGR234 mutant lacked VLCFA, and this mutant was severely impaired in the ability to form functional nodules with the majority of legumes tested. Our work demonstrates the importance of VLCFA in the NGR234-legume symbiosis and also shows that the necessity of BacA for bacteroid differentiation is restricted to specific legume-Rhizobium interactions. KW - Bacteria KW - Rhizobia PY - 2011 U6 - https://doi.org/10.1128/JB.01260-10 SN - 0021-9193 SN - 1098-5530 VL - 193 IS - 9 SP - 2218 EP - 2228 PB - American Society for Microbiology CY - Washington, DC AN - OPUS4-24267 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 -