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      Exploring microbial diversity in Greenland Ice Sheet supraglacial habitats through culturing-dependent and -independent approaches

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          Abstract

          The microbiome of Greenland Ice Sheet supraglacial habitats is still underinvestigated, and as a result there is a lack of representative genomes from these environments. In this study, we investigated the supraglacial microbiome through a combination of culturing-dependent and -independent approaches. We explored ice, cryoconite, biofilm, and snow biodiversity to answer: (1) how microbial diversity differs between supraglacial habitats, (2) if obtained bacterial genomes reflect dominant community members, and (3) how culturing versus high throughput sequencing changes our observations of microbial diversity in supraglacial habitats. Genomes acquired through metagenomic sequencing (133 high-quality MAGs) and whole genome sequencing (73 bacterial isolates) were compared to the metagenome assemblies to investigate abundance within the total environmental DNA. Isolates obtained in this study were not dominant taxa in the habitat they were sampled from, in contrast to the obtained MAGs. We demonstrate here the advantages of using metagenome SSU rRNA genes to reflect whole-community diversity. Additionally, we demonstrate a proof-of-concept of the application of in situ culturing in a supraglacial setting.

          Abstract

          Various culturing-dependent and -independent methods were used to explore microbial diversity on the Greenland Ice Sheet, highlighting the benefits of combining different approaches to capture genomic diversity.

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          Fast gapped-read alignment with Bowtie 2.

          As the rate of sequencing increases, greater throughput is demanded from read aligners. The full-text minute index is often used to make alignment very fast and memory-efficient, but the approach is ill-suited to finding longer, gapped alignments. Bowtie 2 combines the strengths of the full-text minute index with the flexibility and speed of hardware-accelerated dynamic programming algorithms to achieve a combination of high speed, sensitivity and accuracy.
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            RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies

            Motivation: Phylogenies are increasingly used in all fields of medical and biological research. Moreover, because of the next-generation sequencing revolution, datasets used for conducting phylogenetic analyses grow at an unprecedented pace. RAxML (Randomized Axelerated Maximum Likelihood) is a popular program for phylogenetic analyses of large datasets under maximum likelihood. Since the last RAxML paper in 2006, it has been continuously maintained and extended to accommodate the increasingly growing input datasets and to serve the needs of the user community. Results: I present some of the most notable new features and extensions of RAxML, such as a substantial extension of substitution models and supported data types, the introduction of SSE3, AVX and AVX2 vector intrinsics, techniques for reducing the memory requirements of the code and a plethora of operations for conducting post-analyses on sets of trees. In addition, an up-to-date 50-page user manual covering all new RAxML options is available. Availability and implementation: The code is available under GNU GPL at https://github.com/stamatak/standard-RAxML. Contact: alexandros.stamatakis@h-its.org Supplementary information: Supplementary data are available at Bioinformatics online.
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              The SILVA ribosomal RNA gene database project: improved data processing and web-based tools

              SILVA (from Latin silva, forest, http://www.arb-silva.de) is a comprehensive web resource for up to date, quality-controlled databases of aligned ribosomal RNA (rRNA) gene sequences from the Bacteria, Archaea and Eukaryota domains and supplementary online services. The referred database release 111 (July 2012) contains 3 194 778 small subunit and 288 717 large subunit rRNA gene sequences. Since the initial description of the project, substantial new features have been introduced, including advanced quality control procedures, an improved rRNA gene aligner, online tools for probe and primer evaluation and optimized browsing, searching and downloading on the website. Furthermore, the extensively curated SILVA taxonomy and the new non-redundant SILVA datasets provide an ideal reference for high-throughput classification of data from next-generation sequencing approaches.
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                Author and article information

                Contributors
                Journal
                FEMS Microbiol Ecol
                FEMS Microbiol Ecol
                femsec
                FEMS Microbiology Ecology
                Oxford University Press
                0168-6496
                1574-6941
                November 2023
                03 October 2023
                03 October 2023
                : 99
                : 11
                : fiad119
                Affiliations
                Department of Environmental Science, Aarhus University , Frederiksborgvej 399, 4000 Roskilde, Denmark
                Department of Environmental Science, Aarhus University , Frederiksborgvej 399, 4000 Roskilde, Denmark
                Department of Environmental Science, Aarhus University , Frederiksborgvej 399, 4000 Roskilde, Denmark
                Department of Environmental Science, Aarhus University , Frederiksborgvej 399, 4000 Roskilde, Denmark
                Department of Environmental Science, Aarhus University , Frederiksborgvej 399, 4000 Roskilde, Denmark
                Department of Environmental Science, Aarhus University , Frederiksborgvej 399, 4000 Roskilde, Denmark
                Department of Environmental Science, Aarhus University , Frederiksborgvej 399, 4000 Roskilde, Denmark
                German Research Centre for Geosciences , Helmholtz Centre Potsdam, Telegrafenberg, 14473 Potsdam, Germany
                Department of Earth Sciences, Freie Universität Berlin , Malteserstr. 74-100, 12249 Berlin, Germany
                Department of Environmental Science, Aarhus University , Frederiksborgvej 399, 4000 Roskilde, Denmark
                Department of Environmental Science, Aarhus University , Frederiksborgvej 399, 4000 Roskilde, Denmark
                Author notes
                Corresponding authors. Department of Environmental Science, Aarhus University, Frederiksborgvej 399, 4000 Roskilde, Denmark. E-mails: ahj@ 123456envs.au.dk
                Corresponding authors. ama@ 123456envs.au.dk
                Author information
                https://orcid.org/0000-0002-7949-9080
                https://orcid.org/0000-0002-0536-1126
                https://orcid.org/0000-0002-4706-4023
                https://orcid.org/0000-0002-7235-9026
                https://orcid.org/0000-0003-0799-6894
                https://orcid.org/0000-0002-2796-7137
                https://orcid.org/0000-0001-9972-5578
                https://orcid.org/0000-0003-2071-3094
                https://orcid.org/0000-0003-2990-4014
                Article
                fiad119
                10.1093/femsec/fiad119
                10580271
                37791411
                cff9db4e-0b9d-4a93-8830-3e80482f2f5a
                © The Author(s) 2023. Published by Oxford University Press on behalf of FEMS.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 29 April 2023
                : 22 August 2023
                : 28 September 2023
                : 17 October 2023
                Page count
                Pages: 16
                Funding
                Funded by: European Research Council, DOI 10.13039/100010663;
                Award ID: 856416
                Categories
                Research Article
                AcademicSubjects/SCI01150

                Microbiology & Virology
                amplicons,greenland ice sheet, in situ culturing,isolates,mags,metagenome
                Microbiology & Virology
                amplicons, greenland ice sheet, in situ culturing, isolates, mags, metagenome

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