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      The Treasure Vault Can be Opened: Large-Scale Genome Skimming Works Well Using Herbarium and Silica Gel Dried Material

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          Abstract

          Genome skimming has the potential for generating large data sets for DNA barcoding and wider biodiversity genomic studies, particularly via the assembly and annotation of full chloroplast (cpDNA) and nuclear ribosomal DNA (nrDNA) sequences. We compare the success of genome skims of 2051 herbarium specimens from Norway/Polar regions with 4604 freshly collected, silica gel dried specimens mainly from the European Alps and the Carpathians. Overall, we were able to assemble the full chloroplast genome for 67% of the samples and the full nrDNA cluster for 86%. Average insert length, cover and full cpDNA and rDNA assembly were considerably higher for silica gel dried than herbarium-preserved material. However, complete plastid genomes were still assembled for 54% of herbarium samples compared to 70% of silica dried samples. Moreover, there was comparable recovery of coding genes from both tissue sources (121 for silica gel dried and 118 for herbarium material) and only minor differences in assembly success of standard barcodes between silica dried (89% ITS2, 96% matK and rbcL) and herbarium material (87% ITS2, 98% matK and rbcL). The success rate was > 90% for all three markers in 1034 of 1036 genera in 160 families, and only Boraginaceae worked poorly, with 7 genera failing. Our study shows that large-scale genome skims are feasible and work well across most of the land plant families and genera we tested, independently of material type. It is therefore an efficient method for increasing the availability of plant biodiversity genomic data to support a multitude of downstream applications.

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          A DNA barcode for land plants.

          DNA barcoding involves sequencing a standard region of DNA as a tool for species identification. However, there has been no agreement on which region(s) should be used for barcoding land plants. To provide a community recommendation on a standard plant barcode, we have compared the performance of 7 leading candidate plastid DNA regions (atpF-atpH spacer, matK gene, rbcL gene, rpoB gene, rpoC1 gene, psbK-psbI spacer, and trnH-psbA spacer). Based on assessments of recoverability, sequence quality, and levels of species discrimination, we recommend the 2-locus combination of rbcL+matK as the plant barcode. This core 2-locus barcode will provide a universal framework for the routine use of DNA sequence data to identify specimens and contribute toward the discovery of overlooked species of land plants.
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            Increasing phylogenetic resolution at low taxonomic levels using massively parallel sequencing of chloroplast genomes

            Background Molecular evolutionary studies share the common goal of elucidating historical relationships, and the common challenge of adequately sampling taxa and characters. Particularly at low taxonomic levels, recent divergence, rapid radiations, and conservative genome evolution yield limited sequence variation, and dense taxon sampling is often desirable. Recent advances in massively parallel sequencing make it possible to rapidly obtain large amounts of sequence data, and multiplexing makes extensive sampling of megabase sequences feasible. Is it possible to efficiently apply massively parallel sequencing to increase phylogenetic resolution at low taxonomic levels? Results We reconstruct the infrageneric phylogeny of Pinus from 37 nearly-complete chloroplast genomes (average 109 kilobases each of an approximately 120 kilobase genome) generated using multiplexed massively parallel sequencing. 30/33 ingroup nodes resolved with ≥ 95% bootstrap support; this is a substantial improvement relative to prior studies, and shows massively parallel sequencing-based strategies can produce sufficient high quality sequence to reach support levels originally proposed for the phylogenetic bootstrap. Resampling simulations show that at least the entire plastome is necessary to fully resolve Pinus, particularly in rapidly radiating clades. Meta-analysis of 99 published infrageneric phylogenies shows that whole plastome analysis should provide similar gains across a range of plant genera. A disproportionate amount of phylogenetic information resides in two loci (ycf1, ycf2), highlighting their unusual evolutionary properties. Conclusion Plastome sequencing is now an efficient option for increasing phylogenetic resolution at lower taxonomic levels in plant phylogenetic and population genetic analyses. With continuing improvements in sequencing capacity, the strategies herein should revolutionize efforts requiring dense taxon and character sampling, such as phylogeographic analyses and species-level DNA barcoding.
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              The Value of Museum Collections for Research and Society

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                Author and article information

                Journal
                Plants (Basel)
                Plants (Basel)
                plants
                Plants
                MDPI
                2223-7747
                01 April 2020
                April 2020
                : 9
                : 4
                : 432
                Affiliations
                [1 ]Tromsø Museum, UiT—The Arctic University of Norway, N-9037 Tromsø, Norway; marie.f.merkel@ 123456uit.no (M.K.F.M.); youri.lammers@ 123456uit.no (Y.L.); iva.h.pitelkova@ 123456uit.no (I.P.)
                [2 ]LECA, Univ. Grenoble Alpes, Univ. Savoie Mont Blanc, CNRS, F-38000 Grenoble, France; sebastien.lavergne@ 123456univ-grenoble-alpes.fr (S.L.); mboleda1@ 123456xtec.cat (M.B.); charles.pouchon@ 123456univ-grenoble-alpes.fr (C.P.); cristina.roquet@ 123456uab.cat (C.R.); wilfried.thuiller@ 123456univ-grenoble-alpes.fr (W.T.)
                [3 ]Génomique Métabolique, Genoscope, Institut François Jacob, CEA, CNRS, Univ Evry, Université Paris-Saclay, 91057 Evry, France; aalberti@ 123456genoscope.cns.fr (A.A.); fdenoeud@ 123456genoscope.cns.fr (F.D.)
                [4 ]A. Borza Botanical Garden and Faculty of Biology and Geology, Babeș-Bolyai University, 400015 Cluj-Napoca, Romania; mihai.puscas@ 123456ubbcluj.ro
                [5 ]Systematics and Evolution of Vascular Plants (UAB)—Associated Unit to CSIC, Departament de Biologia Animal, Biologia Vegetal i Ecologia, Facultat de Biociències, Universitat Autònoma de Barcelona, ES-08193 Bellaterra, Spain
                [6 ]Institute of Biological Research, National Institute of Research and Development for Biological Sciences, 48 Republicii Street, 400015 Cluj-Napoca, Romania; bogdan.hurdu@ 123456icbcluj.ro
                [7 ]Swiss Federal Research Institute WSL, 8903 Birmensdorf, Switzerland; niklaus.zimmermann@ 123456wsl.ch
                [8 ]Royal Botanic Garden Edinburgh, Edinburgh EH3 5LR, UK; PHollingsworth@ 123456rbge.org.uk
                Author notes
                Author information
                https://orcid.org/0000-0002-8610-1085
                https://orcid.org/0000-0003-3372-9423
                https://orcid.org/0000-0001-8748-3743
                https://orcid.org/0000-0002-3165-108X
                https://orcid.org/0000-0002-5388-5274
                https://orcid.org/0000-0003-3099-9604
                Article
                plants-09-00432
                10.3390/plants9040432
                7238428
                32244605
                e67cee96-eab7-45d5-853d-dd27ceb03ec6
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 27 February 2020
                : 25 March 2020
                Categories
                Article

                alpine,chloroplast dna,environmental dna,its,matk,nuclear ribosomal dna,plant dna barcode,phylogenomic,polar,rbcl

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