@article{GarciaBuckMcMahonetal.2015, author = {Garcia, Sarahi L. and Buck, Moritz and McMahon, Katherine D. and Grossart, Hans-Peter and Eiler, Alexander and Warnecke, Falk}, title = {Auxotrophy and intrapopulation complementary in the "interactome' of a cultivated freshwater model community}, series = {Molecular ecology}, volume = {24}, journal = {Molecular ecology}, number = {17}, publisher = {Wiley-Blackwell}, address = {Hoboken}, issn = {0962-1083}, doi = {10.1111/mec.13319}, pages = {4449 -- 4459}, year = {2015}, abstract = {Microorganisms are usually studied either in highly complex natural communities or in isolation as monoclonal model populations that we manage to grow in the laboratory. Here, we uncover the biology of some of the most common and yet-uncultured bacteria in freshwater environments using a mixed culture from Lake Grosse Fuchskuhle. From a single shotgun metagenome of a freshwater mixed culture of low complexity, we recovered four high-quality metagenome-assembled genomes (MAGs) for metabolic reconstruction. This analysis revealed the metabolic interconnectedness and niche partitioning of these naturally dominant bacteria. In particular, vitamin- and amino acid biosynthetic pathways were distributed unequally with a member of Crenarchaeota most likely being the sole producer of vitamin B12 in the mixed culture. Using coverage-based partitioning of the genes recovered from a single MAG intrapopulation metabolic complementarity was revealed pointing to social' interactions for the common good of populations dominating freshwater plankton. As such, our MAGs highlight the power of mixed cultures to extract naturally occurring interactomes' and to overcome our inability to isolate and grow the microbes dominating in nature.}, language = {en} } @article{GhylinGarciaMoyaetal.2014, author = {Ghylin, Trevor W. and Garcia, Sarahi L. and Moya, Francisco and Oyserman, Ben O. and Schwientek, Patrick and Forest, Katrina T. and Mutschler, James and Dwulit-Smith, Jeffrey and Chan, Leong-Keat and Martinez-Garcia, Manuel and Sczyrba, Alexander and Stepanauskas, Ramunas and Grossart, Hans-Peter and Woyke, Tanja and Warnecke, Falk and Malmstrom, Rex and Bertilsson, Stefan and McMahon, Katherine D.}, title = {Comparative single-cell genomics reveals potential ecological niches for the freshwater acl Actinobacteria lineage}, series = {The ISME journal : multidisciplinary journal of microbial ecology}, volume = {8}, journal = {The ISME journal : multidisciplinary journal of microbial ecology}, number = {12}, publisher = {Nature Publ. Group}, address = {London}, issn = {1751-7362}, doi = {10.1038/ismej.2014.135}, pages = {2503 -- 2516}, year = {2014}, abstract = {Members of the acI lineage of Actinobacteria are the most abundant microorganisms in most freshwater lakes; however, our understanding of the keys to their success and their role in carbon and nutrient cycling in freshwater systems has been hampered by the lack of pure cultures and genomes. We obtained draft genome assemblies from 11 single cells representing three acI tribes (acI-A1, acI-A7, acI-B1) from four temperate lakes in the United States and Europe. Comparative analysis of acI SAGs and other available freshwater bacterial genomes showed that acI has more gene content directed toward carbohydrate acquisition as compared to Polynucleobacter and LD12 Alphaproteobacteria, which seem to specialize more on carboxylic acids. The acI genomes contain actinorhodopsin as well as some genes involved in anaplerotic carbon fixation indicating the capacity to supplement their known heterotrophic lifestyle. Genome-level differences between the acI-A and acI-B clades suggest specialization at the clade level for carbon substrate acquisition. Overall, the acI genomes appear to be highly streamlined versions of Actinobacteria that include some genes allowing it to take advantage of sunlight and N-rich organic compounds such as polyamines, di-and oligopeptides, branched-chain amino acids and cyanophycin. This work significantly expands the known metabolic potential of the cosmopolitan freshwater acI lineage and its ecological and genetic traits.}, language = {en} } @article{SrivastavaMcMahonStepanauskasetal.2016, author = {Srivastava, Abhishek and McMahon, Katherine D. and Stepanauskas, Ramunas and Großart, Hans-Peter}, title = {De novo synthesis and functional analysis of the phosphatase-encoding gene acI-B of uncultured Actinobacteria from Lake Stechlin (NE Germany)}, series = {International microbiology : official journal of the Spanish Society for Microbiology}, volume = {19}, journal = {International microbiology : official journal of the Spanish Society for Microbiology}, publisher = {Institut d'Estudis Catalans}, address = {Barcelona}, issn = {1139-6709}, doi = {10.2436/20.1501.01.262}, pages = {39 -- 47}, year = {2016}, abstract = {The National Center for Biotechnology Information [http://www.ncbi.nlm.nih. gov/guide/taxonomy/] database enlists more than 15,500 bacterial species. But this also includes a plethora of uncultured bacterial representations. Owing to their metabolism, they directly influence biogeochemical cycles, which underscores the the important status of bacteria on our planet. To study the function of a gene from an uncultured bacterium, we have undertaken a de novo gene synthesis approach. Actinobacteria of the acI-B subcluster are important but yet uncultured members of the bacterioplankton in temperate lakes of the northern hemisphere such as oligotrophic Lake Stechlin (NE Germany). This lake is relatively poor in phosphate (P) and harbors on average similar to 1.3 x 10(6) bacterial cells/ml, whereby Actinobacteria of the ac-I lineage can contribute to almost half of the entire bacterial community depending on seasonal variability. Single cell genome analysis of Actinobacterium SCGC AB141-P03, a member of the acI-B tribe in Lake Stechlin has revealed several phosphate-metabolizing genes. The genome of acI-B Actinobacteria indicates potential to degrade polyphosphate compound. To test for this genetic potential, we targeted the exoP-annotated gene potentially encoding polyphosphatase and synthesized it artificially to examine its biochemical role. Heterologous overexpression of the gene in Escherichia coli and protein purification revealed phosphatase activity. Comparative genome analysis suggested that homologs of this gene should be also present in other Actinobacteria of the acI lineages. This strategic retention of specialized genes in their genome provides a metabolic advantage over other members of the aquatic food web in a P-limited ecosystem.}, language = {en} } @article{GrossartMassanaMcMahonetal.2019, author = {Grossart, Hans-Peter and Massana, Ramon and McMahon, Katherine D. and Walsh, David A.}, title = {Linking metagenomics to aquatic microbial ecology and biogeochemical cycles}, series = {Limnology and oceanography}, volume = {65}, journal = {Limnology and oceanography}, publisher = {Wiley}, address = {Hoboken}, issn = {0024-3590}, doi = {10.1002/lno.11382}, pages = {S2 -- S20}, year = {2019}, abstract = {Microbial communities are essential components of aquatic ecosystems through their contribution to food web dynamics and biogeochemical processes. Aquatic microbial diversity is immense and a general challenge is to understand how metabolism and interactions of single organisms shape microbial community dynamics and ecosystem-scale biogeochemical transformations. Metagenomic approaches have developed rapidly, and proven to be powerful in linking microbial community dynamics to biogeochemical processes. In this review, we provide an overview of metagenomic approaches, followed by a discussion on some recent insights they have provided, including those in this special issue. These include the discovery of new taxa and metabolisms in aquatic microbiomes, insights into community assembly and functional ecology as well as evolutionary processes shaping microbial genomes and microbiomes, and the influence of human activities on aquatic microbiomes. Given that metagenomics can now be considered a mature technology where data generation and descriptive analyses are relatively routine and informative, we then discuss metagenomic-enabled research avenues to further link microbial dynamics to biogeochemical processes. These include the integration of metagenomics into well-designed ecological experiments, the use of metagenomics to inform and validate metabolic and biogeochemical models, and the pressing need for ecologically relevant model organisms and simple microbial systems to better interpret the taxonomic and functional information integrated in metagenomes. These research avenues will contribute to a more mechanistic and predictive understanding of links between microbial dynamics and biogeochemical cycles. Owing to rapid climate change and human impacts on aquatic ecosystems, the urgency of such an understanding has never been greater.}, language = {en} } @misc{GarciaBuckHamiltonetal.2018, author = {Garcia, Sarahi L. and Buck, Moritz and Hamilton, Joshua J. and Wurzbacher, Christian and Grossart, Hans-Peter and McMahon, Katherine D. and Eiler, Alexander}, title = {Model communities hint at promiscuous metabolic linkages between ubiquitous free-living freshwater bacteria}, series = {Postprints der Universit{\"a}t Potsdam Mathematisch-Naturwissenschaftliche Reihe}, journal = {Postprints der Universit{\"a}t Potsdam Mathematisch-Naturwissenschaftliche Reihe}, number = {714}, doi = {10.25932/publishup-42729}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-427299}, pages = {8}, year = {2018}, abstract = {Genome streamlining is frequently observed in free-living aquatic microorganisms and results in physiological dependencies between microorganisms. However, we know little about the specificity of these microbial associations. In order to examine the specificity and extent of these associations, we established mixed cultures from three different freshwater environments and analyzed the cooccurrence of organisms using a metagenomic time series. Free-living microorganisms with streamlined genomes lacking multiple biosynthetic pathways showed no clear recurring pattern in their interaction partners. Free-living freshwater bacteria form promiscuous cooperative associations. This notion contrasts with the well-documented high specificities of interaction partners in host-associated bacteria. Considering all data together, we suggest that highly abundant free-living bacterial lineages are functionally versatile in their interactions despite their distinct streamlining tendencies at the single-cell level. This metabolic versatility facilitates interactions with a variable set of community members.}, language = {en} } @article{GarciaBuckHamiltonetal.2018, author = {Garcia, Sarahi L. and Buck, Moritz and Hamilton, Joshua J. and Wurzbacher, Christian and Grossart, Hans-Peter and McMahon, Katherine D. and Eiler, Alexander}, title = {Model communities hint at promiscuous metabolic linkages between ubiquitous free-living freshwater bacteria}, series = {mSphere}, volume = {3}, journal = {mSphere}, number = {3}, publisher = {American Society for Microbiology}, address = {Washington}, issn = {2379-5042}, doi = {10.1128/mSphere.00202-18}, pages = {8}, year = {2018}, abstract = {Genome streamlining is frequently observed in free-living aquatic microorganisms and results in physiological dependencies between microorganisms. However, we know little about the specificity of these microbial associations. In order to examine the specificity and extent of these associations, we established mixed cultures from three different freshwater environments and analyzed the cooccurrence of organisms using a metagenomic time series. Free-living microorganisms with streamlined genomes lacking multiple biosynthetic pathways showed no clear recurring pattern in their interaction partners. Free-living freshwater bacteria form promiscuous cooperative associations. This notion contrasts with the well-documented high specificities of interaction partners in host-associated bacteria. Considering all data together, we suggest that highly abundant free-living bacterial lineages are functionally versatile in their interactions despite their distinct streamlining tendencies at the single-cell level. This metabolic versatility facilitates interactions with a variable set of community members.}, language = {en} } @article{GarciaMcMahonGrossartetal.2014, author = {Garcia, Sarahi L. and McMahon, Katherine D. and Grossart, Hans-Peter and Warnecke, Falk}, title = {Successful enrichment of the ubiquitous freshwater acI Actinobacteria}, series = {Environmental microbiology reports}, volume = {6}, journal = {Environmental microbiology reports}, number = {1}, publisher = {Wiley-Blackwell}, address = {Hoboken}, issn = {1758-2229}, doi = {10.1111/1758-2229.12104}, pages = {21 -- 27}, year = {2014}, abstract = {Actinobacteria of the acI lineage are often the numerically dominant bacterial phylum in surface freshwaters, where they can account for >50\% of total bacteria. Despite their abundance, there are no described isolates. In an effort to obtain enrichment of these ubiquitous freshwater Actinobacteria, diluted freshwater samples from Lake Grosse Fuchskuhle, Germany, were incubated in 96-well culture plates. With this method, a successful enrichment containing high abundances of a member of the lineage acI was established. Phylogenetic classification showed that the acIActinobacteria of the enrichment belonged to the acI-B2 tribe, which seems to prefer acidic lakes. This enrichment grows to low cell densities and thus the oligotrophic nature of acI-B2 was confirmed.}, language = {en} }