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      Low Genetic Differentiation across Three Major Ocean Populations of the Whale Shark, Rhincodon typus

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          Abstract

          Background

          Whale sharks are a declining species for which little biological data is available. While these animals are protected in many parts of their range, they are fished legally and illegally in some countries. Baseline biological and ecological data are needed to allow the formulation of an effective conservation plan for whale sharks. It is not known, for example, whether the whale shark is represented by a single worldwide panmictic population or by numerous, reproductively isolated populations. Genetic analysis of population structure is one essential component of the baseline data required for whale shark conservation.

          Methodology/Principal Findings

          We have identified 8 polymorphic microsatellites in the whale shark and used these markers to assess genetic variation and population structure in a panel of whale sharks covering a broad geographic region. This is the first record of microsatellite loci in the whale shark, which displayed an average of 9 alleles per locus and mean H o = 0.66 and H e = 0.69. All but one of the eight loci meet the expectations of Hardy-Weinberg equilibrium. Analysis of these loci in whale sharks representing three major portions of their range, the Pacific (P), Caribbean (C), and Indian (I) Oceans, determined that there is little population differentiation between animals sampled in different geographic regions, indicating historical gene flow between populations. F ST values for inter-ocean comparisons were low (P×C = 0.0387, C×I = 0.0296 and P×I = −0.0022), and only C×I approached statistical significance (p = 0.0495).

          Conclusions/Significance

          We have shown only low levels of genetic differentiation between geographically distinct whale shark populations. Existing satellite tracking data have revealed both regional and long-range migration of whale sharks throughout their range, which supports the finding of gene flow between populations. Whale sharks traverse geographic and political boundaries during their life history and interbreed with animals from distant populations; conservation efforts must therefore target international protection for this species.

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          Most cited references99

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          Statistical confidence for likelihood-based paternity inference in natural populations.

          Paternity inference using highly polymorphic codominant markers is becoming common in the study of natural populations. However, multiple males are often found to be genetically compatible with each offspring tested, even when the probability of excluding an unrelated male is high. While various methods exist for evaluating the likelihood of paternity of each nonexcluded male, interpreting these likelihoods has hitherto been difficult, and no method takes account of the incomplete sampling and error-prone genetic data typical of large-scale studies of natural systems. We derive likelihood ratios for paternity inference with codominant markers taking account of typing error, and define a statistic delta for resolving paternity. Using allele frequencies from the study population in question, a simulation program generates criteria for delta that permit assignment of paternity to the most likely male with a known level of statistical confidence. The simulation takes account of the number of candidate males, the proportion of males that are sampled and gaps and errors in genetic data. We explore the potentially confounding effect of relatives and show that the method is robust to their presence under commonly encountered conditions. The method is demonstrated using genetic data from the intensively studied red deer (Cervus elaphus) population on the island of Rum, Scotland. The Windows-based computer program, CERVUS, described in this study is available from the authors. CERVUS can be used to calculate allele frequencies, run simulations and perform parentage analysis using data from all types of codominant markers.
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            Modulation of non-templated nucleotide addition by Taq DNA polymerase: primer modifications that facilitate genotyping.

            Taq DNA polymerase can catalyze non-templated addition of a nucleotide (principally adenosine) to the 3' end of PCR-amplified products. Recently, we showed that this activity, which is primer-specific, presents a potential source of error in genotyping studies based on the use of short tandem repeat (STR) markers. Furthermore, in reviewing our data, we found that non-templated nucleotide addition adjacent to a 3' terminal C is favored and that addition adjacent to a 3' terminal A is not. It was clear, however, that features of the template in addition to the 3' terminal base also affect the fraction of product adenylated. To define consensus sequences that promote or inhibit product adenylation, we transplanted sequences between the 5' ends of the reverse primers of markers that are adenylated and those of markers that are not adenylated. It proved difficult to identify a single sequence capable of protecting the products of all markers from non-templated addition of nucleotide. On the other hand, placing the sequence GTTTCTT on the 5' end of reverse primers resulted in nearly 100% adenylation of the 3' end of the forward strand. This modification or related ones (called "PIG-tailing") should facilitate accurate genotyping and efficient T/A cloning.
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              FSTAT (version 1.2): A computer program to calculate F‐statistics

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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2009
                7 April 2009
                : 4
                : 4
                : e4988
                Affiliations
                [1 ]The Department of Biological Sciences, University of Illinois at Chicago, Chicago, Illinois, United States of America
                [2 ]The Shark Research Institute, Princeton, New Jersey, United States of America
                [3 ]The Pritzker Laboratory for Molecular Systematics and Evolution, The Field Museum, Chicago, Illinois, United States of America
                American Museum of Natural History, United States of America
                Author notes

                Conceived and designed the experiments: JVS FO REE MA ML. Performed the experiments: JVS CLS FO KAF. Analyzed the data: JVS FO KAF MA. Contributed reagents/materials/analysis tools: JVS REE KAF ML. Wrote the paper: JVS MA.

                Article
                08-PONE-RA-07706R1
                10.1371/journal.pone.0004988
                2662413
                19352489
                e74f059b-3d6f-42de-a470-21f2b6019add
                Schmidt et al. 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
                : 11 December 2008
                : 7 February 2009
                Page count
                Pages: 9
                Categories
                Research Article
                Ecology/Marine and Freshwater Ecology
                Evolutionary Biology/Animal Genetics
                Genetics and Genomics/Animal Genetics
                Genetics and Genomics/Population Genetics
                Marine and Aquatic Sciences/Genetics, Genomics, and Barcoding

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