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      Evolutionary histories determine DNA barcoding success in vascular plants: seven case studies using intraspecific broad sampling of closely related species

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          Abstract

          Background

          Four plastid regions, rpoB, rpoC1, matK, and trnH- psbA, have been recommended as DNA barcodes for plants. Their success in delimiting species boundaries depends on the existence of a clear-cut difference between inter- and intraspecific variability. We tested the ability of these regions to discriminate among closely related species in seven genera of flowering plants with different generation times (trees, perennials, and annuals). To ensure a maximum coverage of intraspecific diversity, and therefore to better evaluate the resolution power of each barcode, we applied a population genetics approach by sampling three to 45 individuals per species over a wide geographical range.

          Results

          All possible combinations between loci were analysed, which showed that using more than one locus does not always improve the resolution power. The trnH- psbA locus was most effective at discriminating among closely related species ( Acer, Lonicera, Geranium, and Veronica), singly or in combination. For Salix, Adenostyles, and Gentiana, the best results were obtained with the combination of matK, rpoB, and trnH- psbA. No barcoding gap was found within six genera analysed, excepting Lonicera. This is due to shared polymorphisms among species, combined with very divergent sequences within species. These genetic patterns reflect incomplete lineage sorting and hybridization events followed by chloroplast capture.

          Conclusions

          Our results strongly suggest that adding trnH- psbA to the two obligate DNA barcodes proposed by the CBOL plant-working group ( matK and rbcL) should be mandatory for closely related species. In our sampling, generation time had no influence on DNA barcoding success, as the best and worst identification successes were found for the two tree genera ( Acer, 64 % success and Salix, 86 % failure). Evolutionary histories are the main factor influencing DNA barcoding success in the studied genera.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12862-016-0678-0) contains supplementary material, which is available to authorized users.

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          Most cited references46

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          Precision Farming: Technologies and Information as Risk-Reduction Tools

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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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              Use of DNA barcodes to identify flowering plants.

              Methods for identifying species by using short orthologous DNA sequences, known as "DNA barcodes," have been proposed and initiated to facilitate biodiversity studies, identify juveniles, associate sexes, and enhance forensic analyses. The cytochrome c oxidase 1 sequence, which has been found to be widely applicable in animal barcoding, is not appropriate for most species of plants because of a much slower rate of cytochrome c oxidase 1 gene evolution in higher plants than in animals. We therefore propose the nuclear internal transcribed spacer region and the plastid trnH-psbA intergenic spacer as potentially usable DNA regions for applying barcoding to flowering plants. The internal transcribed spacer is the most commonly sequenced locus used in plant phylogenetic investigations at the species level and shows high levels of interspecific divergence. The trnH-psbA spacer, although short ( approximately 450-bp), is the most variable plastid region in angiosperms and is easily amplified across a broad range of land plants. Comparison of the total plastid genomes of tobacco and deadly nightshade enhanced with trials on widely divergent angiosperm taxa, including closely related species in seven plant families and a group of species sampled from a local flora encompassing 50 plant families (for a total of 99 species, 80 genera, and 53 families), suggest that the sequences in this pair of loci have the potential to discriminate among the largest number of plant species for barcoding purposes.
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                Author and article information

                Contributors
                +41 22 418 5165 , +41 22 418 5101 , Yamama.Naciri@ville-ge.ch
                Journal
                BMC Evol Biol
                BMC Evol. Biol
                BMC Evolutionary Biology
                BioMed Central (London )
                1471-2148
                13 May 2016
                13 May 2016
                2016
                : 16
                : 103
                Affiliations
                [ ]Laboratoire de Systématique Végétale et Biodiversité, Conservatoire et Jardin botaniques & University of Geneva, Chemin de l’Impératrice, 1, 1292 Chambésy Geneva, Switzerland
                [ ]SwissBOL, University of Geneva, Department of Genetics and Evolution, Quai Ernest Ansermet 30, 1211 Geneva, Switzerland
                Article
                678
                10.1186/s12862-016-0678-0
                4866073
                27178437
                f8972151-ccdd-45ce-b6b1-438f9d933f8d
                © Caetano Wyler and Naciri. 2016

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 8 January 2016
                : 5 May 2016
                Funding
                Funded by: Swiss National Foundation
                Award ID: 3100A0-120390
                Award Recipient :
                Funded by: Société Académique de Genève
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2016

                Evolutionary Biology
                acer,adenostyles,chloroplast capture,incomplete lineage sorting,interspecific hybridization,gentiana,geranium,lonicera,salix,veronica

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