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      Pangolin Genomes Offer Key Insights and Resources for the World’s Most Trafficked Wild Mammals

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          Abstract

          Pangolins form a group of scaly mammals that are trafficked at record numbers for their meat and purported medicinal properties. Despite their conservation concern, knowledge of their evolution is limited by a paucity of genomic data. We aim to produce exhaustive genomic resources that include 3,238 orthologous genes and whole-genome polymorphisms to assess the evolution of all eight extant pangolin species. Robust orthologous gene-based phylogenies recovered the monophyly of the three genera and highlighted the existence of an undescribed species closely related to Southeast Asian pangolins. Signatures of middle Miocene admixture between an extinct, possibly European, lineage and the ancestor of Southeast Asian pangolins, provide new insights into the early evolutionary history of the group. Demographic trajectories and genome-wide heterozygosity estimates revealed contrasts between continental versus island populations and species lineages, suggesting that conservation planning should consider intraspecific patterns. With the expected loss of genomic diversity from recent, extensive trafficking not yet realized in pangolins, we recommend that populations be genetically surveyed to anticipate any deleterious impact of the illegal trade. Finally, we produce a complete set of genomic resources that will be integral for future conservation management and forensic endeavors for pangolins, including tracing their illegal trade. These comprise the completion of whole-genomes for pangolins through the hybrid assembly of the first reference genome for the giant pangolin ( Smutsia gigantea) and new draft genomes (∼43x–77x) for four additional species, as well as a database of orthologous genes with over 3.4 million polymorphic sites.

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          Trimmomatic: a flexible trimmer for Illumina sequence data

          Motivation: Although many next-generation sequencing (NGS) read preprocessing tools already existed, we could not find any tool or combination of tools that met our requirements in terms of flexibility, correct handling of paired-end data and high performance. We have developed Trimmomatic as a more flexible and efficient preprocessing tool, which could correctly handle paired-end data. Results: The value of NGS read preprocessing is demonstrated for both reference-based and reference-free tasks. Trimmomatic is shown to produce output that is at least competitive with, and in many cases superior to, that produced by other tools, in all scenarios tested. Availability and implementation: Trimmomatic is licensed under GPL V3. It is cross-platform (Java 1.5+ required) and available at http://www.usadellab.org/cms/index.php?page=trimmomatic Contact: usadel@bio1.rwth-aachen.de Supplementary information: Supplementary data are available at Bioinformatics online.
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            The Sequence Alignment/Map format and SAMtools

            Summary: The Sequence Alignment/Map (SAM) format is a generic alignment format for storing read alignments against reference sequences, supporting short and long reads (up to 128 Mbp) produced by different sequencing platforms. It is flexible in style, compact in size, efficient in random access and is the format in which alignments from the 1000 Genomes Project are released. SAMtools implements various utilities for post-processing alignments in the SAM format, such as indexing, variant caller and alignment viewer, and thus provides universal tools for processing read alignments. Availability: http://samtools.sourceforge.net Contact: rd@sanger.ac.uk
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              Fast and accurate short read alignment with Burrows–Wheeler transform

              Motivation: The enormous amount of short reads generated by the new DNA sequencing technologies call for the development of fast and accurate read alignment programs. A first generation of hash table-based methods has been developed, including MAQ, which is accurate, feature rich and fast enough to align short reads from a single individual. However, MAQ does not support gapped alignment for single-end reads, which makes it unsuitable for alignment of longer reads where indels may occur frequently. The speed of MAQ is also a concern when the alignment is scaled up to the resequencing of hundreds of individuals. Results: We implemented Burrows-Wheeler Alignment tool (BWA), a new read alignment package that is based on backward search with Burrows–Wheeler Transform (BWT), to efficiently align short sequencing reads against a large reference sequence such as the human genome, allowing mismatches and gaps. BWA supports both base space reads, e.g. from Illumina sequencing machines, and color space reads from AB SOLiD machines. Evaluations on both simulated and real data suggest that BWA is ∼10–20× faster than MAQ, while achieving similar accuracy. In addition, BWA outputs alignment in the new standard SAM (Sequence Alignment/Map) format. Variant calling and other downstream analyses after the alignment can be achieved with the open source SAMtools software package. Availability: http://maq.sourceforge.net Contact: rd@sanger.ac.uk
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                Author and article information

                Contributors
                Role: Associate Editor
                Journal
                Mol Biol Evol
                Mol Biol Evol
                molbev
                Molecular Biology and Evolution
                Oxford University Press (US )
                0737-4038
                1537-1719
                October 2023
                05 October 2023
                05 October 2023
                : 40
                : 10
                : msad190
                Affiliations
                Laboratoire Evolution et Diversité Biologique (EDB)— IRD-UPS-CNRS, Université Toulouse III , Toulouse, France
                Institut des Sciences de l'Évolution de Montpellier (ISEM), Université de Montpellier, CNRS, IRD , Montpellier, France
                Laboratoire Evolution et Diversité Biologique (EDB)— IRD-UPS-CNRS, Université Toulouse III , Toulouse, France
                Laboratoire Evolution et Diversité Biologique (EDB)— IRD-UPS-CNRS, Université Toulouse III , Toulouse, France
                The State Key Laboratory of Protein and Plant Gene Research of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University , Beijing, China
                Institut des Sciences de l'Évolution de Montpellier (ISEM), Université de Montpellier, CNRS, IRD , Montpellier, France
                Mammal Research Institute, Department of Zoology & Entomology, University of Pretoria , Pretoria, South Africa
                Laboratoire de Parasitologie et Ecologie, Faculté des Sciences, Université de Yaoundé I , Yaoundé, Cameroon
                Mammal Research Institute, Department of Zoology & Entomology, University of Pretoria , Pretoria, South Africa
                Institut des Sciences de l'Évolution de Montpellier (ISEM), Université de Montpellier, CNRS, IRD , Montpellier, France
                The State Key Laboratory of Protein and Plant Gene Research of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University , Beijing, China
                Institut des Sciences de l'Évolution de Montpellier (ISEM), Université de Montpellier, CNRS, IRD , Montpellier, France
                Laboratoire Evolution et Diversité Biologique (EDB)— IRD-UPS-CNRS, Université Toulouse III , Toulouse, France
                CIIMAR/CIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade 16 do Porto, Terminal de Cruzeiros do Porto de Leixões , Porto, Portugal
                Author notes

                Sean P Heighton lead contact.

                Sean P Heighton and Rémi Allio contributed equally to this work.

                Shu-Jin Luo, Frédéric Delsuc and Philippe Gaubert Senior author.

                Conflict of interest statement. The authors declare no competing/conflict of interests.

                Author information
                https://orcid.org/0000-0002-7183-9140
                https://orcid.org/0000-0003-3885-5410
                https://orcid.org/0000-0003-1474-7829
                https://orcid.org/0000-0002-1950-5805
                https://orcid.org/0000-0002-9672-9706
                https://orcid.org/0009-0004-0179-9771
                https://orcid.org/0000-0002-9223-4204
                https://orcid.org/0000-0002-7592-8319
                https://orcid.org/0000-0001-8995-3462
                https://orcid.org/0000-0003-3515-8070
                https://orcid.org/0000-0002-6501-6287
                https://orcid.org/0000-0002-1375-9935
                Article
                msad190
                10.1093/molbev/msad190
                10551234
                37794645
                8b2a61b4-4166-49b5-9b2f-ec09869b969a
                © The Author(s) 2023. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution.

                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
                Page count
                Pages: 18
                Categories
                Discoveries
                AcademicSubjects/SCI01130
                AcademicSubjects/SCI01180

                Molecular biology
                pholidota,full genomes,genomic diversity,conservation,ancient admixture,novel taxon
                Molecular biology
                pholidota, full genomes, genomic diversity, conservation, ancient admixture, novel taxon

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