簡易檢索 / 詳目顯示

研究生: 呂嘉偉
Lu, Chia-Wei
論文名稱: 利用限制酶位點標定之次世代定序技術界定東亞島嶼滑蜥屬物種分化與種化歷史
Species delimitation and speciation history of Scincella spp. of East Asian Islands based on RADseq
指導教授: 林思民
Lin, Si-Min
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2017
畢業學年度: 105
語文別: 英文
論文頁數: 63
中文關鍵詞: 物種界定限制酶位點標定基因滲入滑蜥屬種化歷史
英文關鍵詞: genetic introgression, RAD tags, Scincella, speciation history, species delimitation
DOI URL: https://doi.org/10.6345/NTNU202202806
論文種類: 學術論文
相關次數: 點閱:118下載:6
分享至:
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報
  • 了解物種分化的歷史為現今演化生物學的重要議題,為了探討此議題,分類界線模糊或分化年代相對近的複合群便成為一個合適的研究對象。然而此類研究被認為需要大量基因資料,而使用傳統的定序方法可能無法有效率地取得足夠數量的基因座。近年,次世代定序技術可在短期之內取得大量的基因座與單核苷酸多型性,使我們能更深入地進行更縝密的運算,並執行理論的檢測。滑蜥屬 (Scincella) 是一群外觀極為相似的小型石龍子,物種的界定一直都處於爭議的狀態。例如分布於東亞的寧波滑蜥複合群 (S. modesta complex),成員包含分布於日本的先島滑蜥 (S. boettgeri)、台灣的台灣滑蜥 (S. formosensis) 與中國的寧波滑蜥 (S. modesta);另外尚有數個其他的隱蔽種 (Scincella spp.) 分布於台灣與中國,加深了滑蜥屬物種界定的必要性。我們採用限制酶位點標定定序 (RADseq) 之中的multiplexed shotgun genotyping (MSG) 方法來取得大量的基因座,檢測此複合群的物種界線並重塑其分化歷史。本研究共定序並使用56隻滑蜥做分析,共取得約兩億條短片段序列,並以pyRAD軟體進行序列的分析。隨後使用了367至4188個基因座進行親緣關係、Structure、DAPC與BFD*之物種界定檢測。結果顯示寧波滑蜥複合群包含了六個獨立的支系,其中三個為未被發表過的隱蔽種。另一方面,使用G-PhoCS進行分化歷史的重建,顯示寧波滑蜥複合群主要在上新世晚期至更新世早期分化,且在分化的過程中大部分的物種都有族群數縮減的趨勢。在基因交流檢測方面,我們檢測到了非姊妹種間歷史的基因滲入,顯示了複雜的基因交流歷史。

    Understanding the process and mechanism of speciation has long been a major issue in evolutionary biology. Young species complexes provide excellent opportunities to study the model and process of species diversification. However, the power of evolutionary analyses was limited by the number of available loci. Recently, next-generation sequencing techniques can easily generate numerous loci for testing complex models on how species diverged. In this study, I aim to study the species delimitation of the ground skinks (genus Scincella), a group of small-sized skinks widely distributed throughout North America and Oriental Asia. Due to their morphological similarity, the taxonomic status of Scincella in East Asia is always a controversial issue. Previous studies have shown insufficient diagnosis among S. modesta complex, which comprises S. modesta, S. formosensis, and S. boettgeri. Furthermore, there are several other cryptic taxa (Scincella spp.) recently found in Taiwan and China, which led to taxonomic confusion. In this study, I applied multiplexed shotgun genotyping (MSG) to obtain genome-wide loci. By incorporating coalescent-based methods, I clarified the taxonomic status of members in this species complex and inferred their speciation history. Sequences from 56 individuals were included in analyses, with two hundred million reads reported from 2 reaction lanes. Sequences were assembled and called for single nucleotide polymorphic (SNP) sites using pyRAD. Several analyses (included phylogenetic reconstruction, Structure, DAPC, BFD*) were conducted on 367–4188 RAD loci to show that there are six lineages within S. modesta complex, where 3 of them have not been described before and should be treated as new species. On the other hand, a coalescence-based model, G-PhoCS, revealed that S. modesta complex diverged mainly between middle to late Pleistocene, and most of the taxa experienced population shrinkage. Although there is no evidence of gene flow between sister taxa, historical introgression was detected between non-sister taxa during the process of diversification. The results suggested the complicated history of gene flow among these taxa.

    Introduction.................3 Materials and methods........7 Results.....................16 Discussion..................20 Conclusion..................25 References..................26 Table 1.....................38 Table 2.....................39 Table 3.....................41 Table 4.....................42 Table 5.....................43 Table 6.....................45 Table 7.....................46 Figure 1....................47 Figure 2....................48 Figure 3....................49 Figure 4....................50 Figure 5....................51 Figure 6....................52 Figure 7....................53 Figure 8....................54 Figure 9....................55 Figure 10...................56 Figure 11...................57 Figure 12...................58 Figure 13...................59 Figure 14...................60 Figure 15...................61 Appendix 1..................62 Appendix 2..................63

    Andolfatto P, Davison D, Erezyilmaz D, Hu TT, Mast J, Sunayama-Morita T, Stern DL (2011) Multiplexed shotgun genotyping for rapid and efficient genetic mapping. Genome research 21, 610–617.
    Andrews KR, Good JM, Miller MR, Luikart G, Hohenlohe PA (2016) Harnessing the power of RADseq for ecological and evolutionary genomics. Nature Reviews Genetics 17, 81–92.
    Baird NA, Etter PD, Atwood TS, Currey MC, Shiver AL, Lewis ZA, Selker EU, Cresko WA, Johnson EA (2008) Rapid SNP discovery and mapping using sequced RAD makers. PLoS ONE 3, e3376.
    Barley AJ, Monnahan PJ, Thomson RC, Grismer LL, Brown RM (2015) Sun skink landscape genomics: assessing the roles of micro-evolutionary processes in shaping genetic and phenotypic diversity across a heterogeneous and fragmented landscape. Molecular Ecology 24, 1696–1712.
    Barley AJ, White J, Diesmos AC, Brown RM (2013) The challenge of species delimitation at the extremes: diversification without morphological change in philippine sun skinks. Evolution 67, 3556–3572.
    Bouckaert RR, Heled J, Kuhnert D, Vaughan T, Wu C-H, Xie D, Suchard MA, Rambauet A, Drummond AJ (2014) BEAST 2: a software platform for bayesian evolutionary analysis. PLoS Computational Biology 10, e1003537.
    Bouckaert RR (2010) DensiTree: making sense of sets of phylogenetic trees. Bioinformatics 26, 1372–1373.
    Bryant D, Bouckaert R, Felsenstein J, Rosenberg NA, RoyChoudhury A (2012) Inferring species trees directly from biallelic genetic markers: bypassing gene trees in a full coalescent analysis. Molecular Biology and Evolution 29, 1917–1932.
    Buerkle CA, Gompert Z (2013) Population genomics based on low coverage sequencing: how low should we go? Molecular Ecology 22, 3028–3035.
    Campagna L, Gronau I, Silveira LF, Siepel A, Lovette IJ (2015) Distinguishing noise from signal in patterns of genomic divergence in a highly polymorphic avian radiation. Molecular Ecology 24, 4238–4251.
    Carling MD, Brumfield RT (2007) Gene sampling strategies for multi-locus population estimates of genetic diversity (theta). PLoS ONE 2, e160.
    Carstens BC, Pelletier TA, Reid NM, Satler JD (2013) How to fail at species delimitation. Molecular Ecology 22, 4369–4383.
    Catchen JM, Hohenlohe PA, Bassham S, Amores A, Cresko WA (2013) Stacks: an analysis tool set for population genomics. Molecular Ecology 22, 3124–3140.
    Catchen JM, Amores A, Hohenlohe P, Cresko W, Postlethwait JH (2011) Stacks: building and genotyping loci de novo from short-read sequences. G3: Genes, Genomes, Genetics 1, 171–182.
    Chattopadhyay B, Garg KM, Ramakrishnan U (2014) Effect of diversity and missing data on genetic assignment with RAD-Seq markers. BMC Research Notes 7, 841.
    Chen SL, Ota H, Hikida T (2001) Geographic variation in the two smooth skinks, Scincella boettgeri and S. formosensis (Squamata: Scincidae), in the subtropical East Asian islands. Zoological Science 18, 115–130.
    Cruaud A, Gautier M, Galan M, Foucaud J, Saune L, Genson G, Dubois E, Nidelet S, Deuve T, Rasplus JY (2014) Empirical assessment of RAD sequencing for interspecific phylogeny. Molecular Biology and Evolution 31, 1272–1274.
    Darwin C (1859) On the Origin of Species by Means of Natural Selection, Or, The Preservation of Favoured Races in the Struggle for Life John Murray, London.
    Davey JW, Hohenlohe PA, Etter PD, Boone JQ, Catchen JM, Blaxter ML (2011) Genome-wide genetic marker discovery and genotyping using next-generation sequencing. Nature Reviews Genetics 12, 499–510.
    de Queiroz K (1998) The general lineage concept of species, species crieria, and the process of speciation – a conceptual unification and terminological recommendations. In: Endless Forms: Species and Speciation (eds. Howard SJ, Berlocher SH), pp. 55–78. Oxford University Press, New York.
    de Queiroz K (2005) A unified concept of species and its consequences for the future of taxonomy. Proceedings of the California Academy of Sciences 56, 196.
    de Queiroz K (2007) Species concepts and species delimitation. Systematic Biology 56, 879–886.
    de Queiroz K (2011) Branches in the lines of descent: Charles Darwin and the evolution of the species concept. Biological Journal of the Linnean Society 103, 19–35.
    Earl DA, vonHoldt BM (2012) STRUCTURE HARVESTER: a website and program for visualizing STRUCTURE output and implementing the Evanno method. Conservation Genetics Resources 4, 359–361.
    Eaton DAR (2014) PyRAD: assembly of de novo RADseq loci for phylogenetic analyses. Bioinformatics 30, 1844–1849.
    Eaton DAR, Ree RH (2013) Inferring phylogeny and introgression using RADseq data: an example from flowering plants (Pedicularis: Orobanchaceae). Systematic Biology 62, 689–706.
    Elshire RJ, Glaubitz JC, Sun Q, Poland JA, Kawamoto K, Buckler ES, Mitchell SE (2011) A robust, simple genotyping-by-sequencing (GBS) approach for high diversity species. PLoS ONE 6, e19379.
    Emerson KJ, Merz CR, Catchen JM, Hohenlohe PA, Cresko WA, Bradshaw WE, Holzapfel CM (2010) Resolving postglacial phylogeography using high-throughput sequencing. Proceedings of the National Academy of Sciences of the United States of America 107, 16196–16200.
    Evanno G, Regnaut S, Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Molecular Ecology 14, 2611–2620.
    Ferchaud AL, Hansen MM (2016) The impact of selection, gene flow and demographic history on heterogeneous genomic divergence: three-spine sticklebacks in divergent environments. Molecular Ecology 25, 238–259.
    Fujita MK, Leaché AD, Burbrink FT, McGuire JA, Moritz C (2012) Coalescent-based species delimitation in an integrative taxonomy. Trends in Ecology and Evolution 27, 480–488.
    Glor RE, Gifford ME, Larson A, Losos JB, Schettino LR, Chamizo Lara AR, Jackman TR (2004) Partial island submergence and speciation in an adaptive radiation: a multilocus analysis of the Cuban green anoles. Proceedings of the Royal Society B: Biological Sciences 271, 2257–2265.
    Glor RE, Losos JB, Larson A (2005) Out of Cuba: overwater dispersal and speciation among lizards in the Anolis carolinensis subgroup. Molecular Ecology 14, 2419–2432.
    Gottscho AD, Wood DA, Vandergast AG, Lemos-Espinal J, Gatesy J, Reeder TW (2017) Lineage diversification of fringe-toed lizards (Phrynosomatidae: Uma notata complex) in the Colorado Desert: Delimiting species in the presence of gene flow. Molecular Phylogenetics and Evolution 106, 103–117.
    Greer AE (1974) The generic relationships of the scincid lizard genus Leiolopisma and its relatives. Australian Journal of Zoology supplementary series 31, 1–67.
    Gronau I, Hubisz MJ, Gulko B, Danko CG, Siepel A (2011) Bayesian inference of ancient human demography from individual genome sequences. Nature Genetics 43, 1031–1034.
    Grummer JA, Calderon-Espinosa ML, Nieto-Montes de Oca A, Smith EN, Mendez-de la Cruz FR, Leache AD (2015) Estimating the temporal and spatial extent of gene flow among sympatric lizard populations (genus Sceloporus) in the southern Mexican highlands. Molecular Ecology 24, 1523–1542.
    Hedges SB (2014) The high-level classification of skinks (Reptilia, Squamata, Scincomorpha). Zootaxa 3765, 317–338.
    Hey J, Nielsen R (2004) Multilocus methods for estimating population sizes, migration rates and divergence time, with applications to the divergence of Drosophila pseudoobscura and D. persimilis. Genetics 167, 747–760.
    Hipp AL, Eaton DAR, Cavender-Bares J, Fitzek E, Nipper R, Manos PS (2014) A framework phylogeny of the American oak clade based on sequenced RAD data. PLoS ONE 9, e93975.
    Hohenlohe PA, Bassham S, Etter PD, Stiffler N, Johnson EA, Cresko WA (2010) Population genomics of parallel adaptation in threespine stickleback using sequenced RAD tags. PLoS Genetics 6, e1000862.
    Jackson ND, Austin CC (2009) The combined effects of rivers and refugia generate extreme cryptic fragmentation within the common ground skink (Scincella lateralis). Evolution 64, 409–428.
    Jackson ND, Austin CC (2012) Inferring the evolutionary history of divergence despite gene flow in a lizard species, Scincella lateralis (Scincidae), composed of cryptic lineages. Biological Journal of the Linnean Society 107.
    Jakobsson M, Rosenberg NA (2007) CLUMPP: a cluster matching and permutation program for dealing with label switching and multimodality in analysis of population structure. Bioinformatics 23, 1801–1806.
    Jombart T, Devillard S, Balloux F (2010) Discriminant analysis of principal components a new method for the analysis of genetically structured populations. BMC Genetics 11, 94.
    Kass RE, Raftery AE (1995) Bayes factors. Journal of the American Statistical Association 90, 773–795.
    Koizumi Y, Ota H, Hikida T (2014) Phylogeography of the two smooth skinks, Scincella boettgeri and S. formosensis (Squamata: Scincidae) in the southern Ryukyus and Taiwan, as inferred from variation in mitochondrial cytochrome b sequences. Zoological Science 31, 228–236.
    Lartillot N, Philippe H (2006) Computing Bayes factors using thermodynamic integration. Systematic Biology 55, 195-207.
    Leaché AD, Fujita MK, Minin VN, Bouckaert RR (2014) Species delimitation using genome-wide SNP data. Systematic Biology 63, 534–542.
    Librado P, Rozas J (2009) DnaSP v5: A software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25, 1451–1452.
    Lin SM, Chen CA, Lue KY (2002) Molecular phylogeny and biogeography of the grass lizards genus Takydromus (Reptilia: Lacertidae) of East Asia. Molecular Phylogenetics and Evolution 22, 276–288.
    Lischer HE, Excoffier L (2012) PGDSpider: an automated data conversion tool for connecting population genetics and genomics programs. Bioinformatics 28, 298–299.
    Macher JN, Rozenberg A, Pauls SU, Tollrian R, Wagner R, Leese F (2015) Assessing the phylogeographic history of the montane caddisfly Thremma gallicum using mitochondrial and restriction-site-associated DNA (RAD) markers. Ecology and Evolution 5, 648–662.
    Makino T, Okamoto T, Kurita K, Hikida T (2016) Phylogeography of Ryukyu short-legged skink, Ateuchosaurus pellopleurus, as inferred from mitochondrial cytochrome b sequences In: The eighth world congress of herpetology, Hanzhou, China.
    Martin CH, Crawford JE, Turner BJ, Simons LH (2016) Diabolical survival in Death Valley: recent pupfish colonization, gene flow and genetic assimilation in the smallest species range on earth. Proceedings of the Royal Society B: Biological Sciences 283.
    Nakamura KU, Uéno SI (1963) Japanese Reptiles and Amphibians in Colour Hoikusha, Osaka, Japan.
    Ota H (1998) Geographic patterns of endemism and speciation in amphibians and reptiles of the Ryukyu Archipelago, Japan, with special reference to their paleogeographical implications. Researches on Population Ecology 40, 189–204.
    Ota H (2003) A new subspecies of the agamid lizard, Japalura polygonata (Hallowell, 1861) (Reptilia: Squamata), from Yonagunijima Island of the Yaeyama group, Ryukyu Archipelago. Current herpetology 22, 61–71.
    Ota H, Miyaguni H, Hikida T (1999) Geographic variation in the endemic skink, Ateuchosaurus pellopleurus from the Ryukyu Archipelago, Japan. Journal of Herpetology 33, 106–118.
    Ouboter PE (1986) A revision of the genus Scincella (Reptilia: Sauria: Scincidae) of Asia, with some notes on its evolution. Zoologische Verhandelingen 229, 1–66.
    Osozawa S, Shinjo R, Armid A, Watanabe Y, Horiguchi T, Wakabayashi J (2012) Palaeogeographic reconstructtion of the 1.55Ma synchronous isolation of the Ryukyu Islands Japan, and Taiwan and inflow of the Kuroshio warm current. Interational Geology Review 54, 1369–1388.
    Pante E, Abdelkrim J, Viricel A, Gey D, France SC, Boisselier MC, Samadi S (2015) Use of RAD sequencing for delimiting species. Heredity 114, 450–459.
    Peterson BK, Weber JN, Kay EH, Fisher HS, Hoekstra HE (2012) Double digest RADseq: an inexpensive method for de novo SNP discovery and genotyping in model and non-model species. PLoS ONE 7, e37135.
    Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155, 945 – 959.
    Rambaut A, Drummond AJ (2009) FigTree v1. 3.1. http://tree.bio.ed.ac.uk/software/figtree/
    Rambaut A, Suchard MA, Xie D, Drummond AJ (2014) Tracer v1.6. http://beast.bio.ed.ac.uk/Tracer
    Razkin O, Sonet G, Breugelmans K, Madeira MJ, Gomez-Moliner BJ, Backeljau T (2016) Species limits, interspecific hybridization and phylogeny in the cryptic land snail complex Pyramidula: The power of RADseq data. Molecular Phylogenetics and Evolution 101, 267–278.
    Rittmeyer EN, Austin CC (2015) Combined next-generation sequencing and morphology reveal fine-scale speciation in crocodile skinks (Squamata: Scincidae: Tribolonotus). Molecular Ecology 24, 466–483.
    Rosenberg NA (2003) Distruct: a program for the graphical display of population structure. Molecular Ecology Notes 4, 137–138.
    Rosenberg NA, Nordborg M (2002) Genealogical trees, coalescent theory and the analysis of genetic polymorphisms. Nature Reviews Genetics 3, 380–390.
    Rowe HC, Renaut S, Guggisberg A (2011) RAD in the realm of next-generation sequencing technologies Molecular Ecology 20, 3499–3502.
    Seehausen O, Butlin RK, Keller I, Wagner CE, Boughman JW, Hohenlohe PA, Peichel CL, Saetre GP, Bank C, Brannstrom A, Brelsford A, Clarkson CS, Eroukhmanoff F, Feder JL, Fischer MC, Foote AD, Franchini P, Jiggins CD, Jones FC, Lindholm AK, Lucek K, Maan ME, Marques DA, Martin SH, Matthews B, Meier JI, Most M, Nachman MW, Nonaka E, Rennison DJ, Schwarzer J, Watson ET, Westram AM, Widmer A (2014) Genomics and the origin of species. Nature Reviews Genetics 15, 176–192.
    Sousa V, Hey J (2013) Understanding the origin of species with genome-scale data: modelling gene flow. Nature Reviews Genetics 14, 404–414.
    Stamatakis A (2006) Phylogenetic models of rate heterogeneity: a high performance computing perspective. In: Proceedings of the 20th IEEE/ACM International Parallel and Distributed Processing Symposium.
    Stamatakis A (2014) RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics.
    Streicher JW, Devitt TJ, Goldberg CS, Malone JH, Blackmon H, Fujita MK (2014) Diversification and asymmetrical gene flow across time and space: lineage sorting and hybridization in polytypic barking frogs. Molecular Ecology 23, 3273–3291.
    Tominaga A, Matsui M, Eto K, Ota H (2015) Phylogeny and differentiation of wide-ranging Ryukyu Kajika frog Buergeria japonica (Amphibia: Rhacophoridae): geographic genetic pattern not aimply explained by vicariance through strait formation. Zoological Science 32, 240–247.
    Tseng SP, Li SH, Hsieh CH, Wang HY, Lin SM (2014) Influence of gene flow on divergence dating – implications for the speciation history of Takydromus grass lizards. Molecular Ecology 23, 4770–4784.
    Tseng SP, Wang CJ, Li SH, Lin SM (2015) Within-island speciation with an exceptional case of distinct separation between two sibling lizard species divided by a narrow stream. Molecular Phylogenetics and Evolution 90, 164–175.
    Van Denburgh J (1912) Concerning certain species of reptiles and amphibians from China, Japan, the Loo Choo islands, and Formosa. Proceedings of the California Academy of Sciences 3, 187–258.
    vonHoldt BM, Cahill JA, Fan Z, Gronau I, Robinson J, Pollinger JP, Shapiro B, Wall J, Wayne RK (2016) Whole-genome sequence analysis shows that two endemic species of North American wolf are admixtures of the coyote and gray wolf. Science Advances 2.
    Wang JC (2011) Taxonomy of Scincella and Sphenomorphus (Squamata: Scincidae) of Taiwan based on molecular data and morphological evidence. In: Master's thesis. National Taiwan Normal Unversity, Taiwan, Taipei.
    Winger BM, Hosner PA, Bravo GA, Cuervo AM, Aristizabal N, Cueto LE, Bates JM (2015) Inferring speciation history in the Andes with reduced-representation sequence data: an example in the bay-backed antpittas (Aves; Grallariidae; Grallaria hypoleuca s. l.). Molecular Ecology 24, 6256–6277.
    Wu CI (2001) The genic view of the process of speciation. Journal of Evolutionary Biology 14, 851–865.
    Zarza E, Faircloth BC, Tsai WL, Bryson RW, Jr., Klicka J, McCormack JE (2016) Hidden histories of gene flow in highland birds revealed with genomic markers. Molecular Ecology 25, 5144–5157.

    下載圖示
    QR CODE