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      Delineating Species with DNA Barcodes: A Case of Taxon Dependent Method Performance in Moths

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          Abstract

          The accelerating loss of biodiversity has created a need for more effective ways to discover species. Novel algorithmic approaches for analyzing sequence data combined with rapidly expanding DNA barcode libraries provide a potential solution. While several analytical methods are available for the delineation of operational taxonomic units (OTUs), few studies have compared their performance. This study compares the performance of one morphology-based and four DNA-based (BIN, parsimony networks, ABGD, GMYC) methods on two groups of gelechioid moths. It examines 92 species of Finnish Gelechiinae and 103 species of Australian Elachistinae which were delineated by traditional taxonomy. The results reveal a striking difference in performance between the two taxa with all four DNA-based methods. OTU counts in the Elachistinae showed a wider range and a relatively low (ca. 65%) OTU match with reference species while OTU counts were more congruent and performance was higher (ca. 90%) in the Gelechiinae. Performance rose when only monophyletic species were compared, but the taxon-dependence remained. None of the DNA-based methods produced a correct match with non-monophyletic species, but singletons were handled well. A simulated test of morphospecies-grouping performed very poorly in revealing taxon diversity in these small, dull-colored moths. Despite the strong performance of analyses based on DNA barcodes, species delineated using single-locus mtDNA data are best viewed as OTUs that require validation by subsequent integrative taxonomic work.

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          Bayesian species delimitation using multilocus sequence data.

          In the absence of recent admixture between species, bipartitions of individuals in gene trees that are shared across loci can potentially be used to infer the presence of two or more species. This approach to species delimitation via molecular sequence data has been constrained by the fact that genealogies for individual loci are often poorly resolved and that ancestral lineage sorting, hybridization, and other population genetic processes can lead to discordant gene trees. Here we use a Bayesian modeling approach to generate the posterior probabilities of species assignments taking account of uncertainties due to unknown gene trees and the ancestral coalescent process. For tractability, we rely on a user-specified guide tree to avoid integrating over all possible species delimitations. The statistical performance of the method is examined using simulations, and the method is illustrated by analyzing sequence data from rotifers, fence lizards, and human populations.
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            A cladistic analysis of phenotypic associations with haplotypes inferred from restriction endonuclease mapping and DNA sequence data. III. Cladogram estimation.

            We previously developed a cladistic approach to identify subsets of haplotypes defined by restriction endonuclease mapping or DNA sequencing that are associated with significant phenotypic deviations. Our approach was limited to segments of DNA in which little recombination occurs. In such cases, a cladogram can be constructed from the restriction site or sequence data that represents the evolutionary steps that interrelate the observed haplotypes. The cladogram is used to define a nested statistical design to identify mutational steps associated with significant phenotypic deviations. The central assumption behind this strategy is that any undetected mutation causing a phenotypic effect is embedded within the same evolutionary history that is represented by the cladogram. The power of this approach depends upon the confidence one has in the particular cladogram used to draw inferences. In this paper, we present a strategy for estimating the set of cladograms that are consistent with a particular sample of either restriction site or nucleotide sequence data and that includes the possibility of recombination. We first evaluate the limits of parsimony in constructing cladograms. Once these limits have been determined, we construct the set of parsimonious and nonparsimonious cladograms that is consistent with these limits. Our estimation procedure also identifies haplotypes that are candidates for being products of recombination. If recombination is extensive, our algorithm subdivides the DNA region into two or more subsections, each having little or no internal recombination. We apply this estimation procedure to three data sets to illustrate varying degrees of cladogram ambiguity and recombination.
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              DNA barcodes distinguish species of tropical Lepidoptera.

              Although central to much biological research, the identification of species is often difficult. The use of DNA barcodes, short DNA sequences from a standardized region of the genome, has recently been proposed as a tool to facilitate species identification and discovery. However, the effectiveness of DNA barcoding for identifying specimens in species-rich tropical biotas is unknown. Here we show that cytochrome c oxidase I DNA barcodes effectively discriminate among species in three Lepidoptera families from Area de Conservación Guanacaste in northwestern Costa Rica. We found that 97.9% of the 521 species recognized by prior taxonomic work possess distinctive cytochrome c oxidase I barcodes and that the few instances of interspecific sequence overlap involve very similar species. We also found two or more barcode clusters within each of 13 supposedly single species. Covariation between these clusters and morphological and/or ecological traits indicates overlooked species complexes. If these results are general, DNA barcoding will significantly aid species identification and discovery in tropical settings.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                7 April 2015
                2015
                : 10
                : 4
                : e0122481
                Affiliations
                [1 ]Finnish Museum of Natural History, University of Helsinki, Zoology Unit, University of Helsinki, Helsinki, Finland
                [2 ]Biodiversity Institute of Ontario, University of Guelph, Guelph, Ontario, Canada
                [3 ]Department of Genetics and Physiology, University of Oulu, Oulu, Finland
                [4 ]Metapopulation Research Centre, Department of Biosciences, University of Helsinki, Helsinki, Finland
                University of Veterinary Medicine Hanover, GERMANY
                Author notes

                Competing Interests: HOK-Elanto has been a sponsor for Finnish Museum of Natural History, sponsoring without specifications activities of the institute. It was an internal decision in the Finnish Museum of Natural History to support the publications in the form of financing a collecting trip of Dr. Lauri Kaila to Australia, with no communication with the sponsor regarding this decision. Therefore, the authors declare impartiality and no influence by HOK-Elanto regarding the support for the activity relating to this publication. This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.

                Conceived and designed the experiments: MK. Performed the experiments: MK MN. Analyzed the data: MK MN. Contributed reagents/materials/analysis tools: PDNH MM LK. Wrote the paper: MK MM LK MN PDNH.

                Article
                PONE-D-14-43974
                10.1371/journal.pone.0122481
                4406103
                25849083
                468e39e2-2403-4286-bd62-d9bd7503ea37
                Copyright @ 2015

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 30 September 2014
                : 22 February 2015
                Page count
                Figures: 9, Tables: 3, Pages: 32
                Funding
                This work was supported by the Research Foundation of the University of Helsinki (MK) ( http://www.helsinki.fi/tiedesaatio/english/), Finnish Concordia Fund (MK) ( https://www.konkordia-liitto.com/index.html), Ella and Georg Ehrnrooth Foundation (MK, MN) ( http://www.ellageorg.fi/en/about), Academy of Finland LK (project 1110906) ( http://www.aka.fi/en-GB/A/), HOK-Elanto LK ( http://www.hok-elanto.fi/in-brief/), and the government of Canada through Genome Canada and the Ontario Genomics Institute in support of the International Barcode of Life project PDNH ( http://www.genomecanada.ca/en/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Custom metadata
                All DNA sequences are available from the BOLD database (dataset DS-GELEELA, DOI: 10.5883/DS-GELEELA) and the GenBank database (accession numbers are available in Table S1).

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