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1,003 reference genomes of bacterial and archaeal isolates expand coverage of the tree of life (2017)
Mukherjee, Supratim ; Seshadri, Rekha ; Varghese, Neha J. ; Eloe-Fadrosh, Emiley A. ; Meier-Kolthoff, Jan P. ; Göker, Markus ; Coates, R. Cameron ; Hadjithomas, Michalis ; Pavlopoulos, Georgios A. ; Paez-Espino, David ; Yoshikuni, Yasuo ; Visel, Axel ; Whitman, William B. ; Garrity, George M. ; Eisen, Jonathan A. ; Hugenholtz, Philip ; Pati, Amrita ; Ivanova, Natalia N. ; Woyke, Tanja ; Klenk, Hans-Peter ; Kyrpides, Nikos C.
We present 1,003 reference genomes that were sequenced as part of the Genomic Encyclopedia of Bacteria and Archaea (GEBA) initiative, selected to maximize sequence coverage of phylogenetic space. These genomes double the number of existing type strains and expand their overall phylogenetic diversity by 25%. Comparative analyses with previously available finished and draft genomes reveal a 10.5% increase in novel protein families as a function of phylogenetic diversity. The GEBA genomes recruit 25 million previously unassigned metagenomic proteins from 4,650 samples, improving their phylogenetic and functional interpretation. We identify numerous biosynthetic clusters and experimentally validate a divergent phenazine cluster with potential new chemical structure and antimicrobial activity. This Resource is the largest single release of reference genomes to date. Bacterial and archaeal isolate sequence space is still far from saturated, and future endeavors in this direction will continue to be a valuable resource for scientific discovery.
Phylogenomics of Rhodobacteraceae reveals evolutionary adaptation to marine and non-marine habitats (2017)
Simon, Meinhard ; Scheuner, Carmen ; Meier-Kolthoff, Jan P. ; Brinkhoff, Thorsten ; Wagner-Döbler, Irene ; Ulbrich, Marcus ; Klenk, Hans-Peter ; Schomburg, Dietmar ; Petersen, Jörn ; Göker, Markus
Marine Rhodobacteraceae (Alphaproteobacteria) are key players of biogeochemical cycling, comprise up to 30% of bacterial communities in pelagic environments and are often mutualists of eukaryotes. As ‘Roseobacter clade’, these ‘roseobacters’ are assumed to be monophyletic, but non-marine Rhodobacteraceae have not yet been included in phylogenomic analyses. Therefore, we analysed 106 genome sequences, particularly emphasizing gene sampling and its effect on phylogenetic stability, and investigated relationships between marine versus non-marine habitat, evolutionary origin and genomic adaptations. Our analyses, providing no unequivocal evidence for the monophyly of roseobacters, indicate several shifts between marine and non-marine habitats that occurred independently and were accompanied by characteristic changes in genomic content of orthologs, enzymes and metabolic pathways. Non-marine Rhodobacteraceae gained high-affinity transporters to cope with much lower sulphate concentrations and lost genes related to the reduced sodium chloride and organohalogen concentrations in their habitats. Marine Rhodobacteraceae gained genes required for fucoidan desulphonation and synthesis of the plant hormone indole 3-acetic acid and the compatible solutes ectoin and carnitin. However, neither plasmid composition, even though typical for the family, nor the degree of oligotrophy shows a systematic difference between marine and non-marine Rhodobacteraceae. We suggest the operational term ‘Roseobacter group’ for the marine Rhodobacteraceae strains.
Genome-based taxonomic classification of Bacteroidetes (2016)
Hahnke, Richard L. ; Meier-Kolthoff, Jan P. ; García-López, Marina ; Mukherjee, Supratim ; Huntemann, Marcel ; Ivanova, Natalia N. ; Woyke, Tanja ; Kyrpides, Nikos C. ; Klenk, Hans-Peter ; Göker, Markus
The bacterial phylum Bacteroidetes, characterized by a distinct gliding motility, occurs in a broad variety of ecosystems, habitats, life styles, and physiologies. Accordingly, taxonomic classification of the phylum, based on a limited number of features, proved difficult and controversial in the past, for example, when decisions were based on unresolved phylogenetic trees of the 16S rRNA gene sequence. Here we use a large collection of type-strain genomes from Bacteroidetes and closely related phyla for assessing their taxonomy based on the principles of phylogenetic classification and trees inferred from genome-scale data. No significant conflict between 16S rRNA gene and whole-genome phylogenetic analysis is found, whereas many but not all of the involved taxa are supported as monophyletic groups, particularly in the genome-scale trees. Phenotypic and phylogenomic features support the separation of Balneolaceae as new phylum Balneolaeota from Rhodothermaeota and of Saprospiraceae as new class Saprospiria from Chitinophagia. Epilithonimonas is nested within the older genus Chryseobacterium and without significant phenotypic differences; thus merging the two genera is proposed. Similarly, Vitellibacter is proposed to be included in Aequorivita. Flexibacter is confirmed as being heterogeneous and dissected, yielding six distinct genera. Hallella seregens is a later heterotypic synonym of Prevotella dentalis. Compared to values directly calculated from genome sequences, the G+C content mentioned in many species descriptions is too imprecise; moreover, corrected G+C content values have a significantly better fit to the phylogeny. Corresponding emendations of species descriptions are provided where necessary. Whereas most observed conflict with the current classification of Bacteroidetes is already visible in 16S rRNA gene trees, as expected whole-genome phylogenies are much better resolved.
Proposal of a type strain for Frankia alni (Woronin 1866) Von Tubeuf 1895, emended description of Frankia alni, and recognition of Frankia casuarinae sp. nov. and Frankia elaeagni sp. nov. (2016)
Nouioui, Imen ; Ghodhbane-Gtari, Faten ; Montero-Calasanz, Maria del Carmen ; Göker, Markus ; Meier-Kolthoff, Jan P. ; Schumann, Peter ; Rohde, Manfred ; Goodfellow, Michael ; Fernandez, Maria P. ; Normand, Philippe ; Tisa, Louis S. ; Klenk, Hans-Peter ; Gtari, Maher
Comparative genomics of biotechnologically important yeasts (2016)
Riley, Robert ; Haridas, Sajeet ; Wolfe, Kenneth H. ; Lopes, Mariana R. ; Hittinger, Chris Todd ; Göker, Markus ; Salamov, Asaf A. ; Wisecaver, Jennifer H. ; Long, Tanya M. ; Calvey, Christopher H. ; Aerts, Andrea L. ; Barry, Kerrie W. ; Choi, Cindy ; Clum, Alicia ; Coughlan, Aisling Y. ; Deshpande, Shweta ; Douglass, Alexander P. ; Hanson, Sara J. ; Klenk, Hans-Peter ; LaButti, Kurt M. ; Lapidus, Alla ; Lindquist, Erika A. ; Lipzen, Anna M. ; Meier-Kolthoff, Jan P. ; Ohm, Robin A. ; Otillar, Robert P. ; Pangilinan, Jasmyn L. ; Peng, Yi ; Rokas, Antonis ; Rosa, Carlos A. ; Scheuner, Carmen ; Sibirny, Andriy A. ; Slot, Jason C. ; Stielow, J. Benjamin ; Sun, Hui ; Kurtzman, Cletus P. ; Blackwell, Meredith ; Grigoriev, Igor V. ; Jeffries, Thomas W.
Comparing polysaccharide decomposition between the type strains Gramella echinicola KMM 6050T (DSM 19838T) and Gramella portivictoriae UST040801-001T (DSM 23547T), and emended description of Gramella echinicola Nedashkovskaya et al. 2005 emend. Shahina et al. 2014 and Gramella portivictoriae Lau et al. 2005 (2016)
Panschin, Irina ; Huang, Sixing ; Meier-Kolthoff, Jan P. ; Tindall, Brian J. ; Rohde, Manfred ; Verbarg, Susanne ; Lapidus, Alla ; Han, James ; Trong, Stephan ; Haynes, Matthew ; Reddy, T. B. K. ; Huntemann, Marcel ; Pati, Amrita ; Ivanova, Natalia N. ; Mavromatis, Konstantinos ; Markowitz, Victor ; Woyke, Tanja ; Göker, Markus ; Klenk, Hans-Peter ; Kyrpides, Nikos C. ; Hahnke, Richard L.
Strains of the genus Gramella (family Flavobacteriacae, phylum Bacteroidetes) were isolated from marine habitats such as tidal flat sediments, coastal surface seawater and sea urchins. Flavobacteriaceae have been shown to be involved in the decomposition of plant and algal polysaccharides. However, the potential to decompose polysaccharides may differ tremendously even between species of the same genus. Gramella echinicola KMM 6050T (DSM 19838T) and Gramella portivictoriae UST040801-001T (DSM 23547T) have genomes of similar lengths, similar numbers of protein coding genes and RNA genes. Both genomes encode for a greater number of peptidases compared to ’G. forsetii’. In contrast to the genome of ’G. forsetii’, both genomes comprised a smaller set of CAZymes. Seven polysaccharide utilization loci were identified in the genomes of DSM 19838T and DSM 23547T. Both Gramella strains hydrolyzed starch, galactomannan, arabinoxylan and hydroxyethyl-cellulose, but not pectin, chitosan and cellulose (Avicel). Galactan and xylan were hydrolyzed by strain DSM 19838T, whereas strain DSM 23547T hydrolyzed pachyman and carboxy-methyl cellulose. Conclusively, both Gramella type strains exhibit characteristic physiological, morphological and genomic differences that might be linked to their habitat. Furthermore, the identified enzymes mediating polysaccharide decomposition, are of biotechnological interest.
High-quality draft genome sequence of Flavobacterium suncheonense GH29-5T (DSM 17707T) isolated from greenhouse soil in South Korea, and emended description of Flavobacterium suncheonense GH29-5T (2016)
Tashkandy, Nisreen ; Sabban, Sari ; Fakieh, Mohammad ; Meier-Kolthoff, Jan P. ; Huang, Sixing ; Tindall, Brian J. ; Rohde, Manfred ; Baeshen, Mohammed N. ; Baeshen, Nabih A. ; Lapidus, Alla ; Copeland, Alex ; Pillay, Manoj ; Reddy, T. B. K. ; Huntemann, Marcel ; Pati, Amrita ; Ivanova, Natalia ; Markowitz, Victor ; Woyke, Tanja ; Göker, Markus ; Klenk, Hans-Peter ; Kyrpides, Nikos C. ; Hahnke, Richard L.
Flavobacterium suncheonense is a member of the family Flavobacteriaceae in the phylum Bacteroidetes. Strain GH29-5T (DSM 17707T) was isolated from greenhouse soil in Suncheon, South Korea. F. suncheonense GH29-5T is part of the G enomic E ncyclopedia of B acteria and A rchaea project. The 2,880,663 bp long draft genome consists of 54 scaffolds with 2739 protein-coding genes and 82 RNA genes. The genome of strain GH29-5T has 117 genes encoding peptidases but a small number of genes encoding carbohydrate active enzymes (51 CAZymes). Metallo and serine peptidases were found most frequently. Among CAZymes, eight glycoside hydrolase families, nine glycosyl transferase families, two carbohydrate binding module families and four carbohydrate esterase families were identified. Suprisingly, polysaccharides utilization loci (PULs) were not found in strain GH29-5T. Based on the coherent physiological and genomic characteristics we suggest that F. suncheonense GH29-5T feeds rather on proteins than saccharides and lipids.
GBDP on the grid: a genome-based approach for species delimitation adjusted for an automated and highly parallel processing of large data sets (2014)
Meier-Kolthoff, Jan P. ; Auch, Alexander F. ; Klenk, Hans-Peter ; Göker, Markus
Genome sequence of the chemoheterotrophic soil bacterium Saccharomonospora cyanea type strain (NA-134T) (2013)
Meier-Kolthoff, Jan P. ; Lu, Megan ; Huntemann, Marcel ; Lucas, Susan ; Lapidus, Alla ; Copeland, Alex ; Pitluck, Sam ; Goodwin, Lynne A. ; Han, Cliff ; Tapia, Roxanne ; Pötter, Gabriele ; Land, Miriam ; Ivanova, Natalia ; Rohde, Manfred ; Göker, Markus ; Detter, John C. ; Woyke, Tanja ; Kyrpides, Nikos C. ; Klenk, Hans-Peter
Saccharomonospora cyanea Runmao et al. 1988 is a member of the genus Saccharomonospora in the family Pseudonocardiaceae that is moderately well characterized at the genome level thus far. Members of the genus Saccharomonospora are of interest because they originate from diverse habitats, such as soil, leaf litter, manure, compost, surface of peat, moist, over-heated grain, and ocean sediment, where they probably play a role in the primary degradation of plant material by attacking hemicellulose. Species of the genus Saccharomonospora are usually Gram-positive, non-acid fast, and are classified among the actinomycetes. S. cyanea is characterized by a dark blue (= cyan blue) aerial mycelium. After S. viridis, S. azurea, and S. marina, S. cyanea is only the fourth member in the genus for which a completely sequenced (non-contiguous finished draft status) type strain genome will be published. Here we describe the features of this organism, together with the draft genome sequence, and annotation. The 5,408,301 bp long chromosome with its 5,139 protein-coding and 57 RNA genes was sequenced as part of the DOE funded Community Sequencing Program (CSP) 2010 at the Joint Genome Institute (JGI).
Genotype–phenotype correlations within the Geodermatophilaceae (2022)
Montero-Calasanz, Maria del Carmen ; Yaramis, Adnan ; Rohde, Manfred ; Schumann, Peter ; Klenk, Hans-Peter ; Meier-Kolthoff, Jan P.
The integration of genomic information into microbial systematics along with physiological and chemotaxonomic parameters provides for a reliable classification of prokaryotes. In silico analysis of chemotaxonomic traits is now being introduced to replace characteristics traditionally determined in the laboratory with the dual goal of both increasing the speed of the description of taxa and the accuracy and consistency of taxonomic reports. Genomics has already successfully been applied in the taxonomic rearrangement of Geodermatophilaceae (Actinomycetota) but in the light of new genomic data the taxonomy of the family needs to be revisited. In conjunction with the taxonomic characterisation of four strains phylogenetically located within the family, we conducted a phylogenetic analysis of the whole proteomes of the sequenced type strains and established genotype–phenotype correlations for traits related to chemotaxonomy, cell morphology and metabolism. Results indicated that the four isolates under study represent four novel species within the genus Blastococcus. Additionally, the genera Blastococcus, Geodermatophilus and Modestobacter were shown to be paraphyletic. Consequently, the new genera Trujillonella, Pleomorpha and Goekera were proposed within the Geodermatophilaceae and Blastococcus endophyticus was reclassified as Trujillonella endophytica comb. nov., Geodermatophilus daqingensis as Pleomorpha daqingensis comb. nov. and Modestobacter deserti as Goekera deserti comb. nov. Accordingly, we also proposed emended descriptions of Blastococcus aggregatus, Blastococcus jejuensis, Blastococcus saxobsidens and Blastococcus xanthilyniticus. In silico chemotaxonomic results were overall consistent with wet-lab results. Even though in silico discriminatory levels varied depending on the respective chemotaxonomic trait, this approach is promising for effectively replacing and/or complementing chemotaxonomic analyses at taxonomic ranks above the species level. Finally, interesting but previously overlooked insights regarding morphology and ecology were revealed by the presence of a repertoire of genes related to flagellum synthesis, chemotaxis, spore production and pilus assembly in all representatives of the family. A rich carbon metabolism including four different CO2 fixation pathways and a battery of enzymes able to degrade complex carbohydrates were also identified in Blastococcus genomes.
Taxonomy, physiology, and natural products of Actinobacteria (2016)
Barka, Essaid Ait ; Vatsa, Parul ; Sanchez, Lisa ; Gaveau-Vaillant, Nathalie ; Jacquard, Cedric ; Meier-Kolthoff, Jan P. ; Klenk, Hans-Peter ; Clément, Christophe ; Ouhdouch, Yder ; van Wezel, Gilles P.
Genome-guided analysis allows the identification of novel physiological traits in Trichococcus species (2020)
Strepis, Nikolaos ; Naranjo, Henry D. ; Meier-Kolthoff, Jan P. ; Göker, Markus ; Shapiro, Nicole ; Kyrpides, Nikos ; Klenk, Hans-Peter ; Schaap, Peter J. ; Stams, Alfons J. M. ; Sousa, Diana Z.
Background The genus Trichococcus currently contains nine species: T. flocculiformis, T. pasteurii, T. palustris, T. collinsii, T. patagoniensis, T. ilyis, T. paludicola, T. alkaliphilus, and T. shcherbakoviae. In general, Trichococcus species can degrade a wide range of carbohydrates. However, only T. pasteurii and a non-characterized strain of Trichococcus, strain ES5, have the capacity of converting glycerol to mainly 1,3-propanediol. Comparative genomic analysis of Trichococcus species provides the opportunity to further explore the physiological potential and uncover novel properties of this genus. Results In this study, a genotype-phenotype comparative analysis of Trichococcus strains was performed. The genome of Trichococcus strain ES5 was sequenced and included in the comparison with the other nine type strains. Genes encoding functions related to e.g. the utilization of different carbon sources (glycerol, arabinan and alginate), antibiotic resistance, tolerance to low temperature and osmoregulation could be identified in all the sequences analysed. T. pasteurii and Trichococcus strain ES5 contain a operon with genes encoding necessary enzymes for 1,3-PDO production from glycerol. All the analysed genomes comprise genes encoding for cold shock domains, but only five of the Trichococcus species can grow at 0 °C. Protein domains associated to osmoregulation mechanisms are encoded in the genomes of all Trichococcus species, except in T. palustris, which had a lower resistance to salinity than the other nine studied Trichococcus strains. Conclusions Genome analysis and comparison of ten Trichococcus strains allowed the identification of physiological traits related to substrate utilization and environmental stress resistance (e.g. to cold and salinity). Some substrates were used by single species, e.g. alginate by T. collinsii and arabinan by T. alkaliphilus. Strain ES5 may represent a subspecies of Trichococcus flocculiformis and contrary to the type strain (DSM 2094T), is able to grow on glycerol with the production of 1,3-propanediol.
Genome-based taxonomic classification of the phylum Actinobacteria (2018)
Nouioui, Imen ; Carro, Lorena ; García-López, Marina ; Meier-Kolthoff, Jan P. ; Woyke, Tanja ; Kyrpides, Nikos C. ; Pukall, Rüdiger ; Klenk, Hans-Peter ; Goodfellow, Michael ; Göker, Markus
The application of phylogenetic taxonomic procedures led to improvements in the classification of bacteria assigned to the phylum Actinobacteria but even so there remains a need to further clarify relationships within a taxon that encompasses organisms of agricultural, biotechnological, clinical, and ecological importance. Classification of the morphologically diverse bacteria belonging to this large phylum based on a limited number of features has proved to be difficult, not least when taxonomic decisions rested heavily on interpretation of poorly resolved 16S rRNA gene trees. Here, draft genome sequences of a large collection of actinobacterial type strains were used to infer phylogenetic trees from genome-scale data using principles drawn from phylogenetic systematics. The majority of taxa were found to be monophyletic but several orders, families, and genera, as well as many species and a few subspecies were shown to be in need of revision leading to proposals for the recognition of 2 orders, 10 families, and 17 genera, as well as the transfer of over 100 species to other genera. In addition, emended descriptions are given for many species mainly involving the addition of data on genome size and DNA G+C content, the former can be considered to be a valuable taxonomic marker in actinobacterial systematics. Many of the incongruities detected when the results of the present study were compared with existing classifications had been recognized from 16S rRNA gene trees though whole-genome phylogenies proved to be much better resolved. The few significant incongruities found between 16S/23S rRNA and whole genome trees underline the pitfalls inherent in phylogenies based upon single gene sequences. Similarly good congruence was found between the discontinuous distribution of phenotypic properties and taxa delineated in the phylogenetic trees though diverse non-monophyletic taxa appeared to be based on the use of plesiomorphic character states as diagnostic features.
Genome-based classification of micromonosporae with a focus on their biotechnological and ecological potential (2018)
Carro, Lorena ; Nouioui, Imen ; Sangal, Vartul ; Meier-Kolthoff, Jan P. ; Trujillo, Martha E. ; Montero-Calasanz, Maria del Carmen ; Sahin, Nevzat ; Smith, Darren Lee ; Kim, Kristi E. ; Peluso, Paul ; Deshpande, Shweta ; Woyke, Tanja ; Shapiro, Nicole ; Kyrpides, Nikos C. ; Klenk, Hans-Peter ; Göker, Markus ; Goodfellow, Michael
There is a need to clarify relationships within the actinobacterial genus Micromonospora, the type genus of the family Micromonosporaceae, given its biotechnological and ecological importance. Here, draft genomes of 40 Micromonospora type strains and two non-type strains are made available through the Genomic Encyclopedia of Bacteria and Archaea project and used to generate a phylogenomic tree which showed they could be assigned to well supported phyletic lines that were not evident in corresponding trees based on single and concatenated sequences of conserved genes. DNA G+C ratios derived from genome sequences showed that corresponding data from species descriptions were imprecise. Emended descriptions include precise base composition data and approximate genome sizes of the type strains. antiSMASH analyses of the draft genomes show that micromonosporae have a previously unrealised potential to synthesize novel specialized metabolites. Close to one thousand biosynthetic gene clusters were detected, including NRPS, PKS, terpenes and siderophores clusters that were discontinuously distributed thereby opening up the prospect of prioritising gifted strains for natural product discovery. The distribution of key stress related genes provide an insight into how micromonosporae adapt to key environmental variables. Genes associated with plant interactions highlight the potential use of micromonosporae in agriculture and biotechnology.
Taxonomic use of DNA G+C content and DNA–DNA hybridization in the genomic age (2014)
Meier-Kolthoff, Jan P. ; Klenk, Hans-Peter ; Göker, Markus
Pathways and substrate‐specific regulation of amino acid degradation in Phaeobacter inhibens DSM 17395 (archetype of the marine Roseobacter clade) (2014)
Drüppel, Katharina ; Hensler, Michael ; Trautwein, Kathleen ; Koßmehl, Sebastian ; Wöhlbrand, Lars ; Schmidt‐Hohagen, Kerstin ; Ulbrich, Marcus ; Bergen, Nils ; Meier-Kolthoff, Jan P. ; Göker, Markus ; Klenk, Hans‐Peter ; Schomburg, Dietmar ; Rabus, Ralf
Genome sequence of the moderately thermophilic sulfur-reducing bacterium Thermanaerovibrio velox type strain (Z-9701T) and emended description of the genus Thermanaerovibrio (2013)
Palaniappan, Krishna ; Meier-Kolthoff, Jan P. ; Teshima, Hazuki ; Nolan, Matt ; Lapidus, Alla ; Tice, Hope ; Del Rio, Tijana Glavina ; Cheng, Jan-Fang ; Han, Cliff ; Tapia, Roxanne ; Goodwin, Lynne A. ; Pitluck, Sam ; Liolios, Konstantinos ; Mavromatis, Konstantinos ; Pagani, Ioanna ; Ivanova, Natalia ; Mikhailova, Natalia ; Pati, Amrita ; Chen, Amy ; Rohde, Manfred ; Mayilraj, Shanmugam ; Spring, Stefan ; Detter, John C. ; Göker, Markus ; Bristow, James ; Eisen, Jonathan A. ; Markowitz, Victor ; Hugenholtz, Philip ; Kyrpides, Nikos C. ; Klenk, Hans-Peter ; Woyke, Tanja
Thermanaerovibrio velox Zavarzina et al. 2000 is a member of the Synergistaceae, a family in the phylum Synergistetes that is already well-characterized at the genome level. Members of this phylum were described as Gram-negative staining anaerobic bacteria with a rod/vibrioid cell shape and possessing an atypical outer cell envelope. They inhabit a large variety of anaerobic environments including soil, oil wells, wastewater treatment plants and animal gastrointestinal tracts. They are also found to be linked to sites of human diseases such as cysts, abscesses, and areas of periodontal disease. The moderately thermophilic and organotrophic T. velox shares most of its morphologic and physiologic features with the closely related species, T. acidaminovorans. In addition to Su883T, the type strain of T. acidaminovorans, stain Z-9701T is the second type strain in the genus Thermanaerovibrio to have its genome sequence published. Here we describe the features of this organism, together with the non-contiguous genome sequence and annotation. The 1,880,838 bp long chromosome (non-contiguous finished sequence) with its 1,751 protein-coding and 59 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.
Highly parallelized inference of large genome-based phylogenies (2014)
Meier-Kolthoff, Jan P. ; Auch, Alexander F. ; Klenk, Hans-Peter ; Göker, Markus
When should a DDH experiment be mandatory in microbial taxonomy? (2013)
Meier-Kolthoff, Jan P. ; Göker, Markus ; Spröer, Cathrin ; Klenk, Hans-Peter
Genome sequence-based species delimitation with confidence intervals and improved distance functions (2013)
Meier-Kolthoff, Jan P. ; Auch, Alexander F. ; Klenk, Hans-Peter ; Göker, Markus
Background For the last 25 years species delimitation in prokaryotes (Archaea and Bacteria) was to a large extent based on DNA-DNA hybridization (DDH), a tedious lab procedure designed in the early 1970s that served its purpose astonishingly well in the absence of deciphered genome sequences. With the rapid progress in genome sequencing time has come to directly use the now available and easy to generate genome sequences for delimitation of species. GBDP (Genome Blast Distance Phylogeny) infers genome-to-genome distances between pairs of entirely or partially sequenced genomes, a digital, highly reliable estimator for the relatedness of genomes. Its application as an in-silico replacement for DDH was recently introduced. The main challenge in the implementation of such an application is to produce digital DDH values that must mimic the wet-lab DDH values as close as possible to ensure consistency in the Prokaryotic species concept. Results Correlation and regression analyses were used to determine the best-performing methods and the most influential parameters. GBDP was further enriched with a set of new features such as confidence intervals for intergenomic distances obtained via resampling or via the statistical models for DDH prediction and an additional family of distance functions. As in previous analyses, GBDP obtained the highest agreement with wet-lab DDH among all tested methods, but improved models led to a further increase in the accuracy of DDH prediction. Confidence intervals yielded stable results when inferred from the statistical models, whereas those obtained via resampling showed marked differences between the underlying distance functions. Conclusions Despite the high accuracy of GBDP-based DDH prediction, inferences from limited empirical data are always associated with a certain degree of uncertainty. It is thus crucial to enrich in-silico DDH replacements with confidence-interval estimation, enabling the user to statistically evaluate the outcomes. Such methodological advancements, easily accessible through the web service at http://ggdc.dsmz.de, are crucial steps towards a consistent and truly genome sequence-based classification of microorganisms.
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