1. Syst Appl Microbiol. 2011 May;34(3):169-70. Release LTPs104 of the All-Species Living Tree. Mu... more 1. Syst Appl Microbiol. 2011 May;34(3):169-70. Release LTPs104 of the All-Species Living Tree. Munoz R, Yarza P, Ludwig W, Euzéby J, Amann R, Schleifer KH, Oliver Glöckner F, Rosselló-Móra R. Marine Microbiology Group ...
Marine metatranscriptome data was generated as part of a study investigating the bacterioplankton... more Marine metatranscriptome data was generated as part of a study investigating the bacterioplankton communities towards the end of a diatom-dominated spring phytoplankton bloom. This genomic resource article reports a metatranscriptomic dataset from amidst the winter time prior to the occurrence of the spring diatom bloom. Up to 58% of all sequences could be assigned to predicted genes. Taxonomic analysis based on expressed 16S ribosomal RNA genes identified Alphaproteobacteria and Gammaproteobacteria as the most active community members.
For magnetic orientation, magnetotactic bacteria biosynthesize magnetosomes, which consist of mem... more For magnetic orientation, magnetotactic bacteria biosynthesize magnetosomes, which consist of membrane-enveloped magnetic nanocrystals of either magnetite (Fe3 O4 ) or greigite (Fe3 S4 ). While magnetite formation is increasingly well understood, much less is known about the genetic control of greigite biomineralization. Recently, two related yet distinct sets of magnetosome genes were discovered in a cultivated magnetotactic deltaproteobacterium capable of synthesizing either magnetite or greigite, or both minerals. This led to the conclusion that greigite and magnetite magnetosomes are synthesized by separate biomineralization pathways. Although magnetosomes of both mineral types co-occurred in uncultured multicellular magnetotactic prokaryotes (MMPs), so far only one type of magnetosome genes could be identified in the available genome data. The MMP Candidatus Magnetomorum strain HK-1 from coastal tidal sand flats of the North Sea (Germany) was analysed by a targeted single-cell ...
A recent investigation of bacterioplankton communities in the German Bight towards the end of a d... more A recent investigation of bacterioplankton communities in the German Bight towards the end of a diatom-dominated spring phytoplankton bloom revealed pronounced successions of distinct bacterial clades. A combination of metagenomics and metaproteomics indicated that these clades had distinct substrate spectra and consumed different algal substrates. In this study we re-analyzed samples from the initial study by total community RNA (metatranscriptomics) and 16S rRNA gene amplicon sequencing. This complementary approach provided new insights into the community composition and expressed genes as well as the assessment of metabolic activity levels of distinct clades. Flavobacteria (genera Ulvibacter, Formosa, and Polaribacter), Alphaproteobacteria (SAR11 clade and Rhodobacteraceae) and Gammaproteobacteria (genus Reinekea and SAR92 clade) were the most abundant taxa. Mapping of the metatranscriptome data on assembled and taxonomically classified metagenome data of the same samples substan...
Members of the flavobacterial genus Polaribacter thrive in response to North Sea spring phytoplan... more Members of the flavobacterial genus Polaribacter thrive in response to North Sea spring phytoplankton blooms. We analyzed two respective Polaribacter species by whole genome sequencing, comparative genomics, substrate tests and proteomics. Both can degrade algal polysaccharides but occupy distinct niches. The liquid culture isolate Polaribacter sp. strain Hel1_33_49 has a 3.0-Mbp genome with an overall peptidase:CAZyme ratio of 1.37, four putative polysaccharide utilization loci (PULs) and features proteorhodopsin, whereas the agar plate isolate Polaribacter sp. strain Hel1_85 has a 3.9-Mbp genome with an even peptidase:CAZyme ratio, eight PULs, a mannitol dehydrogenase for decomposing algal mannitol-capped polysaccharides but no proteorhodopsin. Unlike other sequenced Polaribacter species, both isolates have larger sulfatase-rich PULs, supporting earlier assumptions that Polaribacter take part in the decomposition of sulfated polysaccharides. Both strains grow on algal laminarin and the sulfated polysaccharide chondroitin sulfate. For strain Hel1_33_49, we identified by proteomics (i) a laminarin-induced PUL, (ii) chondroitin sulfate-induced CAZymes and (iii) a chondroitin-induced operon that likely enables chondroitin sulfate recognition. These and other data suggest that strain Hel1_33_49 is a planktonic flavobacterium feeding on proteins and a small subset of algal polysaccharides, while the more versatile strain Hel1_85 can decompose a broader spectrum of polysaccharides and likely associates with algae.The ISME Journal advance online publication, 5 December 2014; doi:10.1038/ismej.2014.225.
As an evolutionary marker, 23S ribosomal RNA (rRNA) offers more diagnostic sequence stretches and... more As an evolutionary marker, 23S ribosomal RNA (rRNA) offers more diagnostic sequence stretches and greater sequence variation than 16S rRNA. However, 23S rRNA is still not as widely used. Based on 80 metagenome samples from the Global Ocean Sampling (GOS) Expedition, the usefulness and taxonomic resolution of 23S rRNA were compared to those of 16S rRNA. Since 23S rRNA is approximately twice as large as 16S rRNA, twice as many 23S rRNA gene fragments were retrieved from the GOS reads than 16S rRNA gene fragments, with 23S rRNA gene fragments being generally about 100bp longer. Datasets for 16S and 23S rRNA sequences revealed similar relative abundances for major marine bacterial and archaeal taxa. However, 16S rRNA sequences had a better taxonomic resolution due to their significantly larger reference database. Reevaluation of the specificity of previously published PCR amplification primers and group specific fluorescence in situ hybridization probes on this metagenomic set of non-amplified 23S rRNA sequences revealed that out of 16 primers investigated, only two had more than 90% target group coverage. Evaluations of two probes, BET42a and GAM42a, were in accordance with previous evaluations, with a discrepancy in the target group coverage of the GAM42a probe when evaluated against the GOS metagenomic dataset.
Advances in sequencing technologies challenge the efficient importing and validation of FASTA for... more Advances in sequencing technologies challenge the efficient importing and validation of FASTA formatted sequence data which is still a prerequisite for most bioinformatic tools and pipelines. Comparative analysis of commonly used Bio*-frameworks (BioPerl, BioJava and Biopython) shows that their scalability and accuracy is hampered. FastaValidator represents a platform-independent, standardized, light-weight software library written in the Java programming language. It targets computer scientists and bioinformaticians writing software which needs to parse quickly and accurately large amounts of sequence data. For end-users FastaValidator includes an interactive out-of-the-box validation of FASTA formatted files, as well as a non-interactive mode designed for high-throughput validation in software pipelines. The accuracy and performance of the FastaValidator library qualifies it for large data sets such as those commonly produced by massive parallel (NGS) technologies. It offers scientists a fast, accurate and standardized method for parsing and validating FASTA formatted sequence data.
The co-authors of this paper hereby state their intention to work together to launch the Genomic ... more The co-authors of this paper hereby state their intention to work together to launch the Genomic Observatories Network (GOs Network) for which this document will serve as its Founding Charter. We define a Genomic Observatory as an ecosystem and/or site subject to long-term scientific research, including (but not limited to) the sustained study of genomic biodiversity from single-celled microbes to multicellular organisms.An international group of 64 scientists first published the call for a global network of Genomic Observatories in January 2012. The vision for such a network was expanded in a subsequent paper and developed over a series of meetings in Bremen (Germany), Shenzhen (China), Moorea (French Polynesia), Oxford (UK), Pacific Grove (California, USA), Washington (DC, USA), and London (UK). While this community-building process continues, here we express our mutual intent to establish the GOs Network formally, and to describe our shared vision for its future. The views expres...
Genes involved in magnetite biomineralization are clustered within the genomic magnetosome island... more Genes involved in magnetite biomineralization are clustered within the genomic magnetosome island of Magnetospirillum gryphiswaldense. Their transcriptional organization and regulation were studied by several approaches. Cotranscription of genes within the mamAB, mamDC, and mms clusters was demonstrated by reverse transcription-PCR (RT-PCR) of intergenic regions, indicating the presence of long polycistronic transcripts extending over more than 16 kb. The transcription start
Proceedings of the First ACM International Conference on Bioinformatics and Computational Biology - BCB '10, 2010
... Wolfgang Hankeln1,2, Pier Luigi Buttigieg1,2, Ivaylo Kostadinov1,2, Renzo Kottmann1, Pelin Yi... more ... Wolfgang Hankeln1,2, Pier Luigi Buttigieg1,2, Ivaylo Kostadinov1,2, Renzo Kottmann1, Pelin Yilmaz1,2, Melissa Beth Duhaime1,2, Frank Oliver ... Hy potheses of the corresponding functions were then generated bas ed on the placement and connectivity of vertices repr esenting ...
Publicly available sequence databases of the small subunit ribosomal RNA gene, also known as 16S ... more Publicly available sequence databases of the small subunit ribosomal RNA gene, also known as 16S rRNA in bacteria and archaea, are growing rapidly, and the number of entries currently exceeds 4 million. However, a unified classification and nomenclature framework for all bacteria and archaea does not yet exist. In this Analysis article, we propose rational taxonomic boundaries for high taxa of bacteria and archaea on the basis of 16S rRNA gene sequence identities and suggest a rationale for the circumscription of uncultured taxa that is compatible with the taxonomy of cultured bacteria and archaea. Our analyses show that only nearly complete 16S rRNA sequences give accurate measures of taxonomic diversity. In addition, our analyses suggest that most of the 16S rRNA sequences of the high taxa will be discovered in environmental surveys by the end of the current decade.
Anaerobic oxidation of methane (AOM) in marine sediments is an important microbial process in the... more Anaerobic oxidation of methane (AOM) in marine sediments is an important microbial process in the global carbon cycle and in control of greenhouse gas emission. The responsible organisms supposedly reverse the reactions of methanogenesis, but cultures providing biochemical proof of this have not been isolated. Here we searched for AOM-associated cell components in microbial mats from anoxic methane seeps in
The term “metagenomics” represents a combination of molecular and bioinformatic tools used to ass... more The term “metagenomics” represents a combination of molecular and bioinformatic tools used to assess the genetic information of a community without prior cultivation of the individual species. It is valuable for the study of microorganisms of which only a minor fraction is yet culturable. The collective genomes present in an environmental sample or in an enrichment of target cells are extracted and subject to sequence-based or functional analyses. The field of metagenomics is evolving very rapidly, especially due to newly developed high-throughput sequencing technologies and increased computational power. Metatranscriptome and – proteome analyses are increasingly combined with metagenomic studies in order to assess not only the genetic potential of a microbial community, but also the genes expressed in a particular environment. The present chapter gives a short historical overview of the early years of metagenome analyses, and of possible applications. Challenges regarding the molec...
1. Syst Appl Microbiol. 2011 May;34(3):169-70. Release LTPs104 of the All-Species Living Tree. Mu... more 1. Syst Appl Microbiol. 2011 May;34(3):169-70. Release LTPs104 of the All-Species Living Tree. Munoz R, Yarza P, Ludwig W, Euzéby J, Amann R, Schleifer KH, Oliver Glöckner F, Rosselló-Móra R. Marine Microbiology Group ...
Marine metatranscriptome data was generated as part of a study investigating the bacterioplankton... more Marine metatranscriptome data was generated as part of a study investigating the bacterioplankton communities towards the end of a diatom-dominated spring phytoplankton bloom. This genomic resource article reports a metatranscriptomic dataset from amidst the winter time prior to the occurrence of the spring diatom bloom. Up to 58% of all sequences could be assigned to predicted genes. Taxonomic analysis based on expressed 16S ribosomal RNA genes identified Alphaproteobacteria and Gammaproteobacteria as the most active community members.
For magnetic orientation, magnetotactic bacteria biosynthesize magnetosomes, which consist of mem... more For magnetic orientation, magnetotactic bacteria biosynthesize magnetosomes, which consist of membrane-enveloped magnetic nanocrystals of either magnetite (Fe3 O4 ) or greigite (Fe3 S4 ). While magnetite formation is increasingly well understood, much less is known about the genetic control of greigite biomineralization. Recently, two related yet distinct sets of magnetosome genes were discovered in a cultivated magnetotactic deltaproteobacterium capable of synthesizing either magnetite or greigite, or both minerals. This led to the conclusion that greigite and magnetite magnetosomes are synthesized by separate biomineralization pathways. Although magnetosomes of both mineral types co-occurred in uncultured multicellular magnetotactic prokaryotes (MMPs), so far only one type of magnetosome genes could be identified in the available genome data. The MMP Candidatus Magnetomorum strain HK-1 from coastal tidal sand flats of the North Sea (Germany) was analysed by a targeted single-cell ...
A recent investigation of bacterioplankton communities in the German Bight towards the end of a d... more A recent investigation of bacterioplankton communities in the German Bight towards the end of a diatom-dominated spring phytoplankton bloom revealed pronounced successions of distinct bacterial clades. A combination of metagenomics and metaproteomics indicated that these clades had distinct substrate spectra and consumed different algal substrates. In this study we re-analyzed samples from the initial study by total community RNA (metatranscriptomics) and 16S rRNA gene amplicon sequencing. This complementary approach provided new insights into the community composition and expressed genes as well as the assessment of metabolic activity levels of distinct clades. Flavobacteria (genera Ulvibacter, Formosa, and Polaribacter), Alphaproteobacteria (SAR11 clade and Rhodobacteraceae) and Gammaproteobacteria (genus Reinekea and SAR92 clade) were the most abundant taxa. Mapping of the metatranscriptome data on assembled and taxonomically classified metagenome data of the same samples substan...
Members of the flavobacterial genus Polaribacter thrive in response to North Sea spring phytoplan... more Members of the flavobacterial genus Polaribacter thrive in response to North Sea spring phytoplankton blooms. We analyzed two respective Polaribacter species by whole genome sequencing, comparative genomics, substrate tests and proteomics. Both can degrade algal polysaccharides but occupy distinct niches. The liquid culture isolate Polaribacter sp. strain Hel1_33_49 has a 3.0-Mbp genome with an overall peptidase:CAZyme ratio of 1.37, four putative polysaccharide utilization loci (PULs) and features proteorhodopsin, whereas the agar plate isolate Polaribacter sp. strain Hel1_85 has a 3.9-Mbp genome with an even peptidase:CAZyme ratio, eight PULs, a mannitol dehydrogenase for decomposing algal mannitol-capped polysaccharides but no proteorhodopsin. Unlike other sequenced Polaribacter species, both isolates have larger sulfatase-rich PULs, supporting earlier assumptions that Polaribacter take part in the decomposition of sulfated polysaccharides. Both strains grow on algal laminarin and the sulfated polysaccharide chondroitin sulfate. For strain Hel1_33_49, we identified by proteomics (i) a laminarin-induced PUL, (ii) chondroitin sulfate-induced CAZymes and (iii) a chondroitin-induced operon that likely enables chondroitin sulfate recognition. These and other data suggest that strain Hel1_33_49 is a planktonic flavobacterium feeding on proteins and a small subset of algal polysaccharides, while the more versatile strain Hel1_85 can decompose a broader spectrum of polysaccharides and likely associates with algae.The ISME Journal advance online publication, 5 December 2014; doi:10.1038/ismej.2014.225.
As an evolutionary marker, 23S ribosomal RNA (rRNA) offers more diagnostic sequence stretches and... more As an evolutionary marker, 23S ribosomal RNA (rRNA) offers more diagnostic sequence stretches and greater sequence variation than 16S rRNA. However, 23S rRNA is still not as widely used. Based on 80 metagenome samples from the Global Ocean Sampling (GOS) Expedition, the usefulness and taxonomic resolution of 23S rRNA were compared to those of 16S rRNA. Since 23S rRNA is approximately twice as large as 16S rRNA, twice as many 23S rRNA gene fragments were retrieved from the GOS reads than 16S rRNA gene fragments, with 23S rRNA gene fragments being generally about 100bp longer. Datasets for 16S and 23S rRNA sequences revealed similar relative abundances for major marine bacterial and archaeal taxa. However, 16S rRNA sequences had a better taxonomic resolution due to their significantly larger reference database. Reevaluation of the specificity of previously published PCR amplification primers and group specific fluorescence in situ hybridization probes on this metagenomic set of non-amplified 23S rRNA sequences revealed that out of 16 primers investigated, only two had more than 90% target group coverage. Evaluations of two probes, BET42a and GAM42a, were in accordance with previous evaluations, with a discrepancy in the target group coverage of the GAM42a probe when evaluated against the GOS metagenomic dataset.
Advances in sequencing technologies challenge the efficient importing and validation of FASTA for... more Advances in sequencing technologies challenge the efficient importing and validation of FASTA formatted sequence data which is still a prerequisite for most bioinformatic tools and pipelines. Comparative analysis of commonly used Bio*-frameworks (BioPerl, BioJava and Biopython) shows that their scalability and accuracy is hampered. FastaValidator represents a platform-independent, standardized, light-weight software library written in the Java programming language. It targets computer scientists and bioinformaticians writing software which needs to parse quickly and accurately large amounts of sequence data. For end-users FastaValidator includes an interactive out-of-the-box validation of FASTA formatted files, as well as a non-interactive mode designed for high-throughput validation in software pipelines. The accuracy and performance of the FastaValidator library qualifies it for large data sets such as those commonly produced by massive parallel (NGS) technologies. It offers scientists a fast, accurate and standardized method for parsing and validating FASTA formatted sequence data.
The co-authors of this paper hereby state their intention to work together to launch the Genomic ... more The co-authors of this paper hereby state their intention to work together to launch the Genomic Observatories Network (GOs Network) for which this document will serve as its Founding Charter. We define a Genomic Observatory as an ecosystem and/or site subject to long-term scientific research, including (but not limited to) the sustained study of genomic biodiversity from single-celled microbes to multicellular organisms.An international group of 64 scientists first published the call for a global network of Genomic Observatories in January 2012. The vision for such a network was expanded in a subsequent paper and developed over a series of meetings in Bremen (Germany), Shenzhen (China), Moorea (French Polynesia), Oxford (UK), Pacific Grove (California, USA), Washington (DC, USA), and London (UK). While this community-building process continues, here we express our mutual intent to establish the GOs Network formally, and to describe our shared vision for its future. The views expres...
Genes involved in magnetite biomineralization are clustered within the genomic magnetosome island... more Genes involved in magnetite biomineralization are clustered within the genomic magnetosome island of Magnetospirillum gryphiswaldense. Their transcriptional organization and regulation were studied by several approaches. Cotranscription of genes within the mamAB, mamDC, and mms clusters was demonstrated by reverse transcription-PCR (RT-PCR) of intergenic regions, indicating the presence of long polycistronic transcripts extending over more than 16 kb. The transcription start
Proceedings of the First ACM International Conference on Bioinformatics and Computational Biology - BCB '10, 2010
... Wolfgang Hankeln1,2, Pier Luigi Buttigieg1,2, Ivaylo Kostadinov1,2, Renzo Kottmann1, Pelin Yi... more ... Wolfgang Hankeln1,2, Pier Luigi Buttigieg1,2, Ivaylo Kostadinov1,2, Renzo Kottmann1, Pelin Yilmaz1,2, Melissa Beth Duhaime1,2, Frank Oliver ... Hy potheses of the corresponding functions were then generated bas ed on the placement and connectivity of vertices repr esenting ...
Publicly available sequence databases of the small subunit ribosomal RNA gene, also known as 16S ... more Publicly available sequence databases of the small subunit ribosomal RNA gene, also known as 16S rRNA in bacteria and archaea, are growing rapidly, and the number of entries currently exceeds 4 million. However, a unified classification and nomenclature framework for all bacteria and archaea does not yet exist. In this Analysis article, we propose rational taxonomic boundaries for high taxa of bacteria and archaea on the basis of 16S rRNA gene sequence identities and suggest a rationale for the circumscription of uncultured taxa that is compatible with the taxonomy of cultured bacteria and archaea. Our analyses show that only nearly complete 16S rRNA sequences give accurate measures of taxonomic diversity. In addition, our analyses suggest that most of the 16S rRNA sequences of the high taxa will be discovered in environmental surveys by the end of the current decade.
Anaerobic oxidation of methane (AOM) in marine sediments is an important microbial process in the... more Anaerobic oxidation of methane (AOM) in marine sediments is an important microbial process in the global carbon cycle and in control of greenhouse gas emission. The responsible organisms supposedly reverse the reactions of methanogenesis, but cultures providing biochemical proof of this have not been isolated. Here we searched for AOM-associated cell components in microbial mats from anoxic methane seeps in
The term “metagenomics” represents a combination of molecular and bioinformatic tools used to ass... more The term “metagenomics” represents a combination of molecular and bioinformatic tools used to assess the genetic information of a community without prior cultivation of the individual species. It is valuable for the study of microorganisms of which only a minor fraction is yet culturable. The collective genomes present in an environmental sample or in an enrichment of target cells are extracted and subject to sequence-based or functional analyses. The field of metagenomics is evolving very rapidly, especially due to newly developed high-throughput sequencing technologies and increased computational power. Metatranscriptome and – proteome analyses are increasingly combined with metagenomic studies in order to assess not only the genetic potential of a microbial community, but also the genes expressed in a particular environment. The present chapter gives a short historical overview of the early years of metagenome analyses, and of possible applications. Challenges regarding the molec...
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