Comunidades microbianas de sedimentos costeros crónicamente contaminados con hidrocarburos en regiones polares y subpolares de ambos hemisferios

A pesar de la gran importancia de los microorganismos ambientales para la provisión de servicios ecosistémicos, su estructura y función permanecen aún poco comprendidas debido a su complejidad y bajo porcentaje de miembros cultivables en laboratorio. El objetivo de este trabajo fue estudiar la estructura y potencial metabólico de las comunidades microbianas de ambientes marino-costeros crónicamente contaminados con hidrocarburos en regiones polares y subpolares.Fil: Espínola, Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaFil: Dionisi, Hebe Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaFil: Lozada, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaIX Jornadas Nacionales de Ciencias del Mar y XVII Coloquio de OceanografíaUhsuaia, Tierra del FuegoArgentinaCentro Austral de Investigaciones CientíficasUniversidad Nacional de Tierra del Fueg

Abundance, dynamics, and biogeographic distribution of seven Polycyclic aromatic hydrocarbon dioxygenase gene variants in coastal sediments of Patagonia

Novel polycyclic aromatic hydrocarbon dioxygenase gene variants were present in abundances similar to or higher than those of phnA1 from Cycloclasticus spp. at a chronically polluted subantarctic coastal marine environment in Patagonia. These novel gene variants were detected over a 6-year time span and were also present in sediments from temperate Patagonian sites.Fil: Marcos, Magalí Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico; ArgentinaFil: Lozada, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico; ArgentinaFil: Di Marzio, Walter Dario. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Luján; ArgentinaFil: Dionisi, Hebe Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico; Argentin

The bacterial community structure of hydrocarbon-polluted marine environments as the basis for the definition of an ecological index of hydrocarbon exposure

The aim of this study was to design a molecular biological tool, using information provided by amplicon pyrosequencing of 16S rRNA genes, that could be suitable for environmental assessment and bioremediation in marine ecosystems. We selected 63 bacterial genera that were previously linked to hydrocarbon biodegradation, representing a minimum sample of the bacterial guild associated with this process. We defined an ecological indicator (ecological index of hydrocarbon exposure, EIHE) using the relative abundance values of these genera obtained by pyrotag analysis. This index reflects the proportion of the bacterial community that is potentially capable of biodegrading hydrocarbons. When the bacterial community structures of intertidal sediments from two sites with different pollution histories were analyzed, 16 of the selected genera (25%) were significantly overrepresented with respect to the pristine site, in at least one of the samples from the polluted site. Although the relative abundances of individual genera associated with hydrocarbon biodegradation were generally low in samples from the polluted site, EIHE values were 4 times higher than those in the pristine sample, with at least 5% of the bacterial community in the sediments being represented by the selected genera. EIHE values were also calculated in other oil-exposed marine sediments as well as in seawater using public datasets from experimental systems and field studies. In all cases, the EIHE was significantly higher in oiled than in unpolluted samples, suggesting that this tool could be used as an estimator of the hydrocarbon-degrading potential of microbial communities.Fil: Lozada, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Nacional Patagónico; ArgentinaFil: Marcos, Magalí Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Nacional Patagónico; ArgentinaFil: Commendatore, Marta G.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Nacional Patagónico; ArgentinaFil: Gil, Mónica N.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Nacional Patagónico; ArgentinaFil: Dionisi, Hebe Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Nacional Patagónico; Argentin

Novel aromatic ring-hydroxylating dioxygenase genes from coastal marine sediments of Patagonia

<p>Abstract</p> <p>Background</p> <p>Polycyclic aromatic hydrocarbons (PAHs), widespread pollutants in the marine environment, can produce adverse effects in marine organisms and can be transferred to humans through seafood. Our knowledge of PAH-degrading bacterial populations in the marine environment is still very limited, and mainly originates from studies of cultured bacteria. In this work, genes coding catabolic enzymes from PAH-biodegradation pathways were characterized in coastal sediments of Patagonia with different levels of PAH contamination.</p> <p>Results</p> <p>Genes encoding for the catalytic alpha subunit of aromatic ring-hydroxylating dioxygenases (ARHDs) were amplified from intertidal sediment samples using two different primer sets. Products were cloned and screened by restriction fragment length polymorphism analysis. Clones representing each restriction pattern were selected in each library for sequencing. A total of 500 clones were screened in 9 gene libraries, and 193 clones were sequenced. Libraries contained one to five different ARHD gene types, and this number was correlated with the number of PAHs found in the samples above the quantification limit (<it>r </it>= 0.834, <it>p </it>< 0.05). Overall, eight different ARHD gene types were detected in the sediments. In five of them, their deduced amino acid sequences formed deeply rooted branches with previously described ARHD peptide sequences, exhibiting less than 70% identity to them. They contain consensus sequences of the Rieske type [2Fe-2S] cluster binding site, suggesting that these gene fragments encode for ARHDs. On the other hand, three gene types were closely related to previously described ARHDs: archetypical <it>nahAc</it>-like genes, <it>phnAc</it>-like genes as identified in <it>Alcaligenes faecalis </it>AFK2, and <it>phnA1</it>-like genes from marine PAH-degraders from the genus <it>Cycloclasticus</it>.</p> <p>Conclusion</p> <p>These results show the presence of hitherto unidentified ARHD genes in this sub-Antarctic marine environment exposed to anthropogenic contamination. This information can be used to study the geographical distribution and ecological significance of bacterial populations carrying these genes, and to design molecular assays to monitor the progress and effectiveness of remediation technologies.</p

Predominancia y alta diversidad de genes asociados a la depuración de formas tóxicas de nitrógeno en metagenomas de sedimentos costeros subantárticos de zonas urbanas

Las comunidades microbianas que habitan en los sedimentos son un componente clave de los ecosistemas costeros, ya que impulsan transformaciones biológicas fundamentales como la descomposición de la materia orgánica (MO) y el ciclado de nutrientes, contribuyendo así a la sustentabilidad del ecosistema. En este trabajo utilizamos una estrategia metagenómica para estudiar el rol de las comunidades microbianas asociadas al ciclo del nitrógeno en sedimentos costeros de zonas urbanas de Bahía Ushuaia.Fil: Calderoli, Priscila Anabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaFil: Espinola, Fernando Julian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaFil: Dionisi, Hebe Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaFil: Lozada, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; Argentina7ma Jornada de BecariosPuerto MadrynArgentinaCentro Nacional Patagónic

Microbial assemblages associated with the invasive kelp Undaria pinnatifida in Patagonian coastal waters: Structure and alginolytic potential

Undaria pinnatifida is a brown algae native to Asia that has settled in various regions worldwide, periodically contributing with large quantities of C and nutrients during its annual cycle. In this work, we analyzed a coastal site in Patagonia (Argentina) that has been colonized for three decades by U. pinnatifida, focusing on associated microbial communities in three different compartments. An important influence of algae was observed in seawater, especially in the bottom of the algal forest during the austral summer (January) at the moment of greater biomass release. This was evidenced by changes in DOC concentration and its quality indicators (higher Freshness and lower Humification index) and higher DIC. Although maximum values of NH4 and PO4 were observed in January, bottom water samples had lower concentrations than surface water, suggesting nutrient consumption by bacteria during algal DOM release. Concomitantly, bacterial abundance peaked, reaching 4.68 ± 1.33 × 105 cells mL −1 (January), showing also higher capability of degrading alginate, a major component of brown algae cell walls. Microbial community structure was influenced by sampling date, season, sampling zone (surface or bottom), and environmental factors (temperature, salinity, pH, dissolved oxygen, nutrients). Samples of epiphytic biofilms showed a distinct community structure compared to seawater, lower diversity, and remarkably high alginolytic capability, suggesting adaptation to degrade algal biomass. A high microdiversity of populations of the genus Leucothrix (Gammaproteobacteria, Thiotrichales) that accounted for a large fraction of epiphytic communities was observed, and changed over time. Epiphytic assemblages shared more taxa with bottom than with surface seawater assemblages, indicating a certain level of exchange between communities in the forest surroundings. This work provides insight into the impact of U. pinnatifida decay on seawater quality, and the role of microbial communities on adapting to massive biomass inputs through rapid DOM turnover.Fil: Lozada, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaFil: Zabala, Maria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biología de Organismos Marinos; ArgentinaFil: Garcia, Patricia Elizabeth. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; ArgentinaFil: Dieguez, Maria del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; ArgentinaFil: Bigatti, Gregorio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biología de Organismos Marinos; ArgentinaFil: Fermani, Paulina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaFil: Unrein, Fernando. Universidad Nacional de San Martin. Instituto Tecnologico de Chascomus. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - la Plata. Instituto Tecnologico de Chascomus.; ArgentinaFil: Dionisi, Hebe Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; Argentin

Prospecting biotechnologically-relevant monooxygenases from cold sediment metagenomes: An in silico approach

Source at https://doi.org/10.3390/md15040114.The goal of this work was to identify sequences encoding monooxygenase biocatalysts with novel features by in silico mining an assembled metagenomic dataset of polar and subpolar marine sediments. The targeted enzyme sequences were Baeyer–Villiger and bacterial cytochrome P450 monooxygenases (CYP153). These enzymes have wide-ranging applications, from the synthesis of steroids, antibiotics, mycotoxins and pheromones to the synthesis of monomers for polymerization and anticancer precursors, due to their extraordinary enantio-, regio-, and chemo- selectivity that are valuable features for organic synthesis. Phylogenetic analyses were used to select the most divergent sequences affiliated to these enzyme families among the 264 putative monooxygenases recovered from the ~14 million protein-coding sequences in the assembled metagenome dataset. Three-dimensional structure modeling and docking analysis suggested features useful in biotechnological applications in five metagenomic sequences, such as wide substrate range, novel substrate specificity or regioselectivity. Further analysis revealed structural features associated with psychrophilic enzymes, such as broader substrate accessibility, larger catalytic pockets or low domain interactions, suggesting that they could be applied in biooxidations at room or low temperatures, saving costs inherent to energy consumption. This work allowed the identification of putative enzyme candidates with promising features from metagenomes, providing a suitable starting point for further developments

Alginate Lyases and Fucanases from Uncultured Microorganisms of Coastal Sediments of Southern Patagonia

Fucoidans and alginates, major polysaccharides of the brown algae cell matrix, have various biological properties. Their high molecular weight hinder some biomedical applications, but this limitation can be overcome through enzymatic degradation to obtain bioactive oligosaccharides. The goal of this work was to identify and characterize fucanases and alginate lyases from uncultured marine bacteria, related to poorly characterized CAZy families (GH107, GH168 and PL38). Using a series of HMMs specific for these families, homologous sequences were identified in a metagenomic dataset from intertidal sediments of Ushuaia Bay (Tierra del Fuego Island, Argentina), a Subantarctic environment with high brown algae biomass. Seven sequences related to the GH107 family (9-40 % protein identity with members of the family) and 21 sequences related to the GH168 family (9 a 80 % identity) were identified in the dataset. In addition, 16 sequences were related to members of the PL38 family. Most members of this family contain a PF05426 (Alginate Lyase) domain, but the only biochemically characterized enzyme is a glucuronan lyase with negligible alginate lyase activity. Putative PL38 sequences showed 30-71 % identity with members of the family. Based on in silico analyses, we selected four sequences related to the GH107 family and two related to the PL38 family for heterologous expression. Fucanase activity was evaluated by carbohydrate polyacrylamide gel electrophoresis (C-PAGE), with four substrates from different species of brown algae. One of the expressed proteins, #113643 (probably from a Planctomycetes), showed the highest level of expression, and the addition of the purified enzyme resulted in the degradation of fucoidans faster than previously characterized members of the GH107 family, in the substrate purified from Macrocystis pyrifera. The enzyme was active at low temperatures (5 - 45 °C), and in a wide range of salinity concentrations (9.5 - 861 mM NaCl) and pH values (4.5 - 9). Alginate lyase activity was measured by following the increase in Abs235nm. Proteins #216008 (Gammaproteobacteria) and #149193 (Bacteroidetes) also showed high expression levels and both enzymes displayed alginate lyase activity, adding a new activity to the PL38 family. This study increases the current knowledge on fucoidan and alginate degradation potential in microbial communities from Subantarctic sediments, and describes novel enzymes from bacteria of an extreme environment for brown algae biomass utilization. JG and CM contributed equally to the work.Fil: Gonzalez, Jessica Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaFil: Molina, Celeste Amancay Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaFil: Musumeci, Matias Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Entre Ríos. Universidad Nacional de Entre Ríos. Centro de Investigaciones y Transferencia de Entre Ríos; ArgentinaFil: Lozada, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biología de Organismos Marinos; ArgentinaFil: Ponce, Nora Marta Andrea. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Ciudad Universitaria. Centro de Investigaciones en Hidratos de Carbono. Subsede del Centro de Investigaciones en Hidratos de Carbono | Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Centro de Investigaciones en Hidratos de Carbono. Subsede del Centro de Investigaciones en Hidratos de Carbono; ArgentinaFil: Stortz, Carlos Arturo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Ciudad Universitaria. Centro de Investigaciones en Hidratos de Carbono. Subsede del Centro de Investigaciones en Hidratos de Carbono | Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Centro de Investigaciones en Hidratos de Carbono. Subsede del Centro de Investigaciones en Hidratos de Carbono; ArgentinaFil: Dionisi, Hebe Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaWorld Microbe ForumWashingtonEstados UnidosAmerican Society for MicrobiologyFederation of European Microbiological Societie

Neutral Lipid Biosynthetic Potential in Sediment Microbial Communities from Subantarctic Environments

Bacteria from a limited number of taxa are known to accumulate wax esters (WE) and triacylglycerol (TAG) as an adaptation response to stressful environmental conditions, although this capability is poorly understood at the microbial community level. The goal of this work was to uncover the prevalence and diversity of bacteria with the potential to synthesize neutral lipids in coastal sediments of Subantarctic and Antarctic environments, and to characterize the gene clusters related to this process. More than 48,000 sequences containing the PF03007 domain (specific of the key enzyme wax ester synthase/acyl-CoA:diacylglycerol acyltransferase, WS/DGAT) were retrieved from 13 metagenomes, including subtidal and intertidal sediments of Ushuaia Bay, Argentina (54° 48’ S, 68° 17’ W), and subtidal sediments of Potter Cove, 25 de Mayo Island, Antarctica (62° 13’ S, 58° 39’ W). Abundance of putative WS/DGAT sequences in the sediment metagenomes was 1.23 ± 0.42 times relative to 12 single-copy genes encoding ribosomal proteins, much higher than in seawater (0.13 ± 0.31 times in 338 metagenomes). In an ordination analysis, the metagenomes were grouped by geographic location, although closely related sequences were present in both environments despite a 1,000 km distance and the potential barrier of the Antarctic Circumpolar Current. Most sequences were binned to the Proteobacteria or the Actinobacteria phyla. Phylogenetic analysis revealed that the majority of the identified sequences were most closely related to sequences from genomes assembled from metagenomes, from environmental samples including seawater, marine sediments, groundwater, freshwater and biological wastewater treatment plants. The genomic context of putative WS/DGAT sequences included genes encoding putative Type-2 PAPs and HAD-type hydrolases, glycerol- and acylglycerol- phosphate O-acyltransferases, some of them potentially responsible for specific steps in WE and TAG biosynthesis. In addition, some scaffolds contained genes of related pathways such as fatty-acids metabolism, suggesting carbon recycling might drive the flux to neutral lipid synthesis. These results indicate the presence of abundant and diverse bacterial populations with the potential to synthesize lipid storage compounds. This information increases our understanding on the mechanisms used by bacteria from extreme environments to adapt to environmental stressors. FP and VG contributed equally.Fil: Pascutti, Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Galvan, Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Sandoval, Natalia Elisa. Universidad Nacional de la Patagonia "San Juan Bosco". Instituto de Biociencias de la Patagonia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biociencias de la Patagonia; ArgentinaFil: Lanfranconi, Mariana Patricia. Universidad Nacional de la Patagonia "San Juan Bosco". Instituto de Biociencias de la Patagonia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biociencias de la Patagonia; ArgentinaFil: Lozada, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biología de Organismos Marinos; ArgentinaFil: Arabolaza, Ana Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Mac Cormack, Walter Patricio. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; ArgentinaFil: Alvarez, Hector Manuel. Universidad Nacional de la Patagonia "San Juan Bosco". Instituto de Biociencias de la Patagonia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biociencias de la Patagonia; ArgentinaFil: Gramajo, Hugo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Dionisi, Hebe Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaWorld Microbe ForumWashingtonEstados UnidosAmerican Society for MicrobiologyFederation of European Microbiological Societie

A communal catalogue reveals Earth's multiscale microbial diversity

Our growing awareness of the microbial world's importance and diversity contrasts starkly with our limited understanding of its fundamental structure. Despite recent advances in DNA sequencing, a lack of standardized protocols and common analytical frameworks impedes comparisons among studies, hindering the development of global inferences about microbial life on Earth. Here we present a meta-analysis of microbial community samples collected by hundreds of researchers for the Earth Microbiome Project. Coordinated protocols and new analytical methods, particularly the use of exact sequences instead of clustered operational taxonomic units, enable bacterial and archaeal ribosomal RNA gene sequences to be followed across multiple studies and allow us to explore patterns of diversity at an unprecedented scale. The result is both a reference database giving global context to DNA sequence data and a framework for incorporating data from future studies, fostering increasingly complete characterization of Earth's microbial diversity.Peer reviewe