TY - JOUR A1 - Garcia, Sarahi L. A1 - Buck, Moritz A1 - McMahon, Katherine D. A1 - Grossart, Hans-Peter A1 - Eiler, Alexander A1 - Warnecke, Falk T1 - Auxotrophy and intrapopulation complementary in the "interactome' of a cultivated freshwater model community JF - Molecular ecology N2 - 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. KW - community KW - cultures KW - interactions KW - metagenomics KW - populations Y1 - 2015 U6 - https://doi.org/10.1111/mec.13319 SN - 0962-1083 SN - 1365-294X VL - 24 IS - 17 SP - 4449 EP - 4459 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Ghylin, Trevor W. A1 - Garcia, Sarahi L. A1 - Moya, Francisco A1 - Oyserman, Ben O. A1 - Schwientek, Patrick A1 - Forest, Katrina T. A1 - Mutschler, James A1 - Dwulit-Smith, Jeffrey A1 - Chan, Leong-Keat A1 - Martinez-Garcia, Manuel A1 - Sczyrba, Alexander A1 - Stepanauskas, Ramunas A1 - Grossart, Hans-Peter A1 - Woyke, Tanja A1 - Warnecke, Falk A1 - Malmstrom, Rex A1 - Bertilsson, Stefan A1 - McMahon, Katherine D. T1 - Comparative single-cell genomics reveals potential ecological niches for the freshwater acl Actinobacteria lineage JF - The ISME journal : multidisciplinary journal of microbial ecology N2 - 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. Y1 - 2014 U6 - https://doi.org/10.1038/ismej.2014.135 SN - 1751-7362 SN - 1751-7370 VL - 8 IS - 12 SP - 2503 EP - 2516 PB - Nature Publ. Group CY - London ER - TY - JOUR A1 - Srivastava, Abhishek A1 - McMahon, Katherine D. A1 - Stepanauskas, Ramunas A1 - Großart, Hans-Peter T1 - De novo synthesis and functional analysis of the phosphatase-encoding gene acI-B of uncultured Actinobacteria from Lake Stechlin (NE Germany) JF - International microbiology : official journal of the Spanish Society for Microbiology N2 - 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. KW - acI-B in Actinobacteria KW - phosphatases KW - single cell genomics KW - phosphate limitation KW - Lake Stechlin KW - NE Germany Y1 - 2016 U6 - https://doi.org/10.2436/20.1501.01.262 SN - 1139-6709 SN - 1618-1905 VL - 19 SP - 39 EP - 47 PB - Institut d'Estudis Catalans CY - Barcelona ER - TY - JOUR A1 - Grossart, Hans-Peter A1 - Massana, Ramon A1 - McMahon, Katherine D. A1 - Walsh, David A. T1 - Linking metagenomics to aquatic microbial ecology and biogeochemical cycles JF - Limnology and oceanography N2 - 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. Y1 - 2019 U6 - https://doi.org/10.1002/lno.11382 SN - 0024-3590 SN - 1939-5590 VL - 65 SP - S2 EP - S20 PB - Wiley CY - Hoboken ER - TY - GEN A1 - Garcia, Sarahi L. A1 - Buck, Moritz A1 - Hamilton, Joshua J. A1 - Wurzbacher, Christian A1 - Grossart, Hans-Peter A1 - McMahon, Katherine D. A1 - Eiler, Alexander T1 - Model communities hint at promiscuous metabolic linkages between ubiquitous free-living freshwater bacteria T2 - Postprints der Universität Potsdam Mathematisch-Naturwissenschaftliche Reihe N2 - 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. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 714 KW - community KW - interactions KW - metagenomics KW - microbial ecology KW - mixed cultures KW - promiscuous Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-427299 IS - 714 ER - TY - JOUR A1 - Garcia, Sarahi L. A1 - Buck, Moritz A1 - Hamilton, Joshua J. A1 - Wurzbacher, Christian A1 - Grossart, Hans-Peter A1 - McMahon, Katherine D. A1 - Eiler, Alexander T1 - Model communities hint at promiscuous metabolic linkages between ubiquitous free-living freshwater bacteria JF - mSphere N2 - 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. KW - community KW - interactions KW - metagenomics KW - microbial ecology KW - mixed cultures KW - promiscuous Y1 - 2018 U6 - https://doi.org/10.1128/mSphere.00202-18 SN - 2379-5042 VL - 3 IS - 3 PB - American Society for Microbiology CY - Washington ER - TY - JOUR A1 - Garcia, Sarahi L. A1 - McMahon, Katherine D. A1 - Grossart, Hans-Peter A1 - Warnecke, Falk T1 - Successful enrichment of the ubiquitous freshwater acI Actinobacteria JF - Environmental microbiology reports N2 - 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. Y1 - 2014 U6 - https://doi.org/10.1111/1758-2229.12104 SN - 1758-2229 VL - 6 IS - 1 SP - 21 EP - 27 PB - Wiley-Blackwell CY - Hoboken ER -