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      Identifying Genetic Signatures of Natural Selection Using Pooled Population Sequencing in Picea abies

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          Abstract

          The joint inference of selection and past demography remain a costly and demanding task. We used next generation sequencing of two pools of 48 Norway spruce mother trees, one corresponding to the Fennoscandian domain, and the other to the Alpine domain, to assess nucleotide polymorphism at 88 nuclear genes. These genes are candidate genes for phenological traits, and most belong to the photoperiod pathway. Estimates of population genetic summary statistics from the pooled data are similar to previous estimates, suggesting that pooled sequencing is reliable. The nonsynonymous SNPs tended to have both lower frequency differences and lower F ST values between the two domains than silent ones. These results suggest the presence of purifying selection. The divergence between the two domains based on synonymous changes was around 5 million yr, a time similar to a recent phylogenetic estimate of 6 million yr, but much larger than earlier estimates based on isozymes. Two approaches, one of them novel and that considers both F ST and difference in allele frequencies between the two domains, were used to identify SNPs potentially under diversifying selection. SNPs from around 20 genes were detected, including genes previously identified as main target for selection, such as PaPRR3 and PaGI.

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          Most cited references28

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          Estimating and interpreting F ST: The impact of rare variants

          In a pair of seminal papers, Sewall Wright and Gustave Malécot introduced F ST as a measure of structure in natural populations. In the decades that followed, a number of papers provided differing definitions, estimation methods, and interpretations beyond Wright's. While this diversity in methods has enabled many studies in genetics, it has also introduced confusion regarding how to estimate F ST from available data. Considering this confusion, wide variation in published estimates of F ST for pairs of HapMap populations is a cause for concern. These estimates changed—in some cases more than twofold—when comparing estimates from genotyping arrays to those from sequence data. Indeed, changes in F ST from sequencing data might be expected due to population genetic factors affecting rare variants. While rare variants do influence the result, we show that this is largely through differences in estimation methods. Correcting for this yields estimates of F ST that are much more concordant between sequence and genotype data. These differences relate to three specific issues: (1) estimating F ST for a single SNP, (2) combining estimates of F ST across multiple SNPs, and (3) selecting the set of SNPs used in the computation. Changes in each of these aspects of estimation may result in F ST estimates that are highly divergent from one another. Here, we clarify these issues and propose solutions.
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            The next generation of molecular markers from massively parallel sequencing of pooled DNA samples.

            Next generation sequencing (NGS) is about to revolutionize genetic analysis. Currently NGS techniques are mainly used to sequence individual genomes. Due to the high sequence coverage required, the costs for population-scale analyses are still too high to allow an extension to nonmodel organisms. Here, we show that NGS of pools of individuals is often more effective in SNP discovery and provides more accurate allele frequency estimates, even when taking sequencing errors into account. We modify the population genetic estimators Tajima's π and Watterson's to obtain unbiased estimates from NGS pooling data. Given the same sequencing effort, the resulting estimators often show a better performance than those obtained from individual sequencing. Although our analysis also shows that NGS of pools of individuals will not be preferable under all circumstances, it provides a cost-effective approach to estimate allele frequencies on a genome-wide scale.
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              Measures of divergence between populations and the effect of forces that reduce variability.

              Wright's FST and related statistics are often used to measure the extent of divergence among populations of the same species relative to the net genetic diversity within the species. This paper compares several definitions of FST which are relevant to DNA sequence data, and shows that these must be used with care when estimating migration parameters. It is also pointed out that FST is strongly influenced by the level of within-population diversity. In situations where factors such as selection on closely linked sites are expected to have stronger effects on within-population diversity at some loci than at others, differences among loci can result entirely from differences in within-population diversities. It is shown that several published cases of differences in FST among regions of high and low recombination in Drosophila may be caused in this way. For the purpose of comparisons of levels of between-population differences among loci or species which are subject to different intensities of forces that reduce variability within local populations, absolute measures of divergence between populations should be used in preference to relative measures such as FST.
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                Author and article information

                Journal
                G3 (Bethesda)
                Genetics
                G3: Genes, Genomes, Genetics
                G3: Genes, Genomes, Genetics
                G3: Genes, Genomes, Genetics
                G3: Genes|Genomes|Genetics
                Genetics Society of America
                2160-1836
                2 May 2016
                July 2016
                : 6
                : 7
                : 1979-1989
                Affiliations
                [* ]Department of Ecology and Genetics, Evolutionary Biology Centre and Science for Life Laboratory, Uppsala University, 75236 Sweden
                []Department of Agricultural, Food, Environmental and Animal Sciences, University of Udine, 33100 Italy
                Author notes
                [1]

                Present address: Department of Medical Biochemistry and Microbiology, Uppsala University, 75236 Sweden.

                [2]

                Present address: Key Laboratory of Stress Physiology and Ecology in Cold and Arid Regions, Chinese Academy of Sciences, Lanzhou, China.

                [3 ]Corresponding author: Uppsala University, Norbyvägen 18 D, 75236 Uppsala, Sweden. E-mail: martin.lascoux@ 123456ebc.uu.se
                Author information
                http://orcid.org/0000-0003-1699-9042
                Article
                GGG_028753
                10.1534/g3.116.028753
                4938651
                27172202
                5860e1c5-053b-47a4-94ec-f2a9e8bf812b
                Copyright © 2016 Chen et al.

                This is an open-access article 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 the original work is properly cited.

                History
                : 05 March 2016
                : 28 April 2016
                Page count
                Figures: 5, Tables: 3, Equations: 2, References: 55, Pages: 11
                Categories
                Investigations

                Genetics
                pooled sequencing,fst,allele frequencies,local adaptation
                Genetics
                pooled sequencing, fst, allele frequencies, local adaptation

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