Abstract
Babina holsti is a ranid frog that occurs on northern Okinawa (Yanbaru) and Tokashiki Islands of southwest Japan. Because of its narrow distribution and recent habitat destruction, its population size is believed to have decreased, and the species has been listed as an endangered species on the IUCN Red List. To promote effective conservation management, we examined the population structure, gene flow, and demographic history of B. holsti using a mitochondrial gene and 12 microsatellite loci. The population structure analyses first distinguished the Tokashiki Island population from the Yanbaru ones. The Yanbaru populations were further divided into main and southernmost populations. Large genetic differences were detected between the Tokashiki and Yanbaru populations, and their divergence time was estimated as 0.38 Mya. Based on demographic analyses on Yanbaru, population expansion generally occurred after the last glacial period and then decreased rapidly. Specifically, only the southern Yanbaru populations have experienced a population decline within the past 5000 years. These results suggested that prehistoric geographic events may have affected the historical population decline on Yanbaru. On Tokashiki, the lowest genetic diversity and a significant population bottleneck were detected, both probably caused by scarcer environmental resources. These results are beneficial to sustainable conservation management of B. holsti, especially the conservation priority of the populations in Tokashiki and southern Yanbaru.





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Birky CW Jr, Maruyama T, Fuerst P (1983) An approach to population and evolutionary theory for genes in mitochondria and chloroplasts, and some results. Genetics 103:513–527
Bryant EH, Backus VL, Clark ME, Reed DH (1999) Experimental tests of captive breeding for endangered species. Conserv Biol 13:1487–1496
Chen X, Chen Z, Jiang J, Qiao L, Lu Y, Zhou K, Zheng G, Zhai X, Liu J (2013) Molecular phylogeny and diversification of the genus Odorrana (Amphibia, Anura, Ranidae) inferred from two mitochondrial genes. Mol Phylogenet Evol 69:1196–1202
Clement M, Posada D, Crandall KA (2000) TCS: a computer program to estimate gene genealogies. Mol Ecol 9:1657–1659
Cornuet JM, Luikart G (1996) Description and power analysis of two tests for detecting recent population bottlenecks from allele frequency data. Genetics 144:2001–2014
Crochet PA (2000) Genetic structure of avian populations—allozymes revisited. Mol Ecol 9:1463–1469
Donoghue PCJ, Benton MJ (2007) Rocks and clocks: calibrating the tree of life using fossils and molecules. Trends Ecol Evol 22:424–431
Drummond AJ, Suchard MA (2010) Bayesian random local clocks, or one rate to rule them all. BMC Biol 8:114–125
Drummond AJ, Rambaut A, Shapiro B, Pybus OG (2005) Bayesian coalescent inference of past population dynamics from molecular sequences. Mol Biol Evol 22:1185–1192
Earl DA, VonHoldt BM (2011) STRUCTURE HARVESTER: a website and program for visualizing STRUCTURE output and implementing the Evanno method. Conserv Genet Res 4:359–361
Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32:1792–1797
Environment Agency of Japan (2000) Threatened wildlife of Japan—red data book, 2nd edn, vol 3. Japan Wildlife Research Center, Tokyo
Evanno G, Regnaut S, Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Mol Ecol 14:2611–2620
Excoffier L, Lischer HEL (2010) Arlequin suite ver. 3.5: a new series of programs to perform population genetics analyses under Linux and Windows. Mol Ecol Resour 10:564–567
Excoffier L, Slatkin M (1998) Incorporating genotypes of relatives into a test of linkage disequilibrium. Am J Hum Genet 62:171–180
Frankham R (1998) Inbreeding and extinction: island populations. Conserv Biol 12:665–675
Frankham R, Ballou JD, Briscoe DA (2002) Introduction to conservation genetics. Cambridge University Press, New York
Fu Y-X (1997) Statistical tests of neutrality of mutations against population growth, hitchhiking and background selection. Genetics 147:915–925
Funk WC, Blouin MS, Corn PS, Maxell BA, Pilliod DS, Amish S, Allendorf FW (2005) Population structure of Columbia spotted frogs (Rana luteiventris) is strongly affected by the landscape. Mol Ecol 14:483–496
Galtier N, Nabholz B, Glémin S, Hurst GDD (2009) Mitochondrial DNA as a marker of molecular diversity: a reappraisal. Mol Ecol 18:4541–4550
Garza JC, Williamson EG (2001) Detection of reduction in population size using data from microsatellite loci. Mol Ecol 10:305–318
Guindon S, Gascuel O (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol 52:696–704
Guo SW, Thompson EA (1992) Performing the exact test of Hardy–Weinberg proportion for multiple alleles. Biometrics 48:361–372
Hewitt GM (2004) The structure of biodiversity—insights from molecular phylogeography. Front Zool 1:4
Hoegg S, Vences M, Brinkmann H, Meyer A (2004) Phylogeny and comparative substitution rates of frogs inferred from sequences of three nuclear genes. Mol Biol Evol 21:1188–1200
Honda M, Matsui M, Tominaga A, Ota H, Tanaka S (2012) Phylogeny and biogeography of the Anderson’s crocodile newt, Echinotriton andersoni (Amphibia: Caudata), as revealed by mitochondrial DNA sequences. Mol Phylogenet Evol 65:642–653
Hongo C, Kayanne H (2010) Holocene sea-level record from corals: reliability of paleodepth indicators at Ishigaki Island, Ryukyu Islands, Japan. Palaeogeogr Palaeoclimatol Palaeoecol 287:143–151
Hubisz MJ, Falush D, Stephens M, Pritchard JK (2009) Inferring weak population structure with the assistance of sample group information. Mol Ecol Resour 9:1322–1332
Igawa T, Kurabayashi A, Usuki C, Fujii T, Sumida M (2008) Complete mitochondrial genomes of three neobatrachian anurans: a case study of divergence time estimation using different data and calibration settings. Gene 407:116–129
Igawa T, Oumi S, Katsuren S, Sumida M (2013) Population structure and landscape genetics of two endangered frog species of genus Odorrana: different scenarios on two islands. Heredity 110:46–56
Iha K (1966) Distribution and eating habits of mongoose in Okinawa. Okinawa Agric 5:39–44
Ihaka R, Gentleman R (1996) R: a language for data analysis and graphics. J Comput Graph Stat 5:299–314
IUCN (2013) IUCN red list of threatened species. Version 2013.2. http://www.iucnredlist.org. Accessed 23 May 2014
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
Jarne P, Lagoda PJL (1996) Microsatellites, from molecules to populations and back. Trends Ecol Evol 11:424–429
Kakehashi R, Igawa T, Iwai N, Shoda-Kagaya E, Sumida M (2013a) Development and characterization of new microsatellite loci in the Otton frog (Babina subaspera) and cross-amplification in a congeneric species, Holst’s frog (B. holsti). Conserv Genet Res 5:1071–1073
Kakehashi R, Kurabayashi A, Oumi S, Katsuren S, Hoso M, Sumida M (2013b) Mitochondrial genomes of Japanese Babina frogs (Ranidae, Anura): unique gene arrangements and the phylogenetic position of genus Babina. Genes Genet Syst 88:59–67
Kan H (2014) Formation process of coral reefs in Ryukyu Islands. In: Takamiya H, Shinzato T (eds) Empirical studies for environmental and cultural change in pre- and proto-historic era, vol 2. Rokuichi Shobo, Tokyo (in Japanese)
Knopp T, Cano JM, Crochet PA, Merilä J (2007) Contrasting levels of variation in neutral and quantitative genetic loci on island populations of moor frogs (Rana arvalis). Conserv Genet 8:45–56
Kuroda T, Ozawa T (1996) Paleoclimatic and vegetational changes during the Pleistocene and Holocene in the Ryukyu Islands inferred from pollen assemblages. J Geogr 105:328–342
Librado P, Rozas J (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25:1451–1452
Lisiecki LE, Raymo ME (2005) A Pliocene–Pleistocene stack of 57 globally distributed benthic δ18 O records. Paleoceanography 20:PA1003
Maeda N, Matsui M (1999) Frogs and toads of Japan. Bun-ichi Sogo Shuppan, Tokyo
Manni F, Guerard E, Heyer E (2004) Geographic patterns of (genetic, morphologic, linguistic) variation: how barriers can be detected by using Monmonier’s algorithm. Hum Biol 76:173–190
Matsui M, Shimada T, Ota H, Tanaka-Ueno T (2005) Multiple invasions of the Ryukyu Archipelago by oriental frogs of the subgenus Odorrana with phylogenetic reassessment of the related subgenera of the genus Rana. Mol Phylogenet Evol 37:733–742
Ministry of the Environment Government of Japan (2012) Small Indian mongoose eradication project in the northern part of Okinawa Island. Naha Nature Conservation Office Web. http://kyushu.env.go.jp/naha/pre_2013/0703b.html. Accessed 23 May 2014
Monmonier MS (1973) Maximum-difference barriers: an alternative numerical regionalization method. Geogr Anal 3:245–261
Nei M, Maruyama T, Chakraborty R (1975) The bottleneck effect and genetic variability in populations. Evolution 29:1–10
Nei M, Tajima F, Tateno Y (1983) Accuracy of estimated phylogenetic trees from molecular data. J Mol Evol 19:153–170
Newman D, Pilson D (1997) Increased probability of extinction due to decreased genetic effective population size: experimental populations of Clarkia pulchella. Evolution 51:354–362
Ogura G, Sasaki T, Toyama M, Takehara K, Nakachi M, Ishibashi O, Kawashima Y, Oda S (2002) Food habitats of the feral small Asian mongoose (Herpestes javanicus) and impacts on native species in the northern part of Okinawa Island. Mamm Sci 41:53–62
Ohta T, Kimura M (1973) A model of mutation appropriate to estimate the number of electrophoretically detectable alleles in a finite population. Genet Res 22:201–204
Omoto K (2004) Radiocarbon ages and isotope fractionations of beachrock samples collected from the Nansei Islands, southwestern Japan. Radiocarbon 46:539–550
Peakall R, Smouse PE (2012) GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research—an update. Bioinformatics 28:2537–2539
Phillips S, Anderson R, Schapire R (2006) Maximum entropy modeling of species geographic distributions. Ecol Model 190:231–259
Piry S, Luikart G, Cornuet J-M (1999) BOTTLENECK: a computer program for detecting recent reductions in the effective population size using allele frequency data. J Hered 90:502–503
Posada D (2008) jModelTest: phylogenetic model averaging. Mol Biol Evol 25:1253–1256
Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155:945–959
Pulliam HR (1988) Sources, sinks, and population regulation. Am Nat 132:652–661
Rambaut A, Drummond AJ (2007) Tracer v1.4. http://beast.bio.ed.ac.uk/Tracer. Accessed 28 May 2014
Raymond M, Rousset F (1995) An exact test for population differentiation. Evolution 49:1280–1283
Rosenberg NA (2004) Distruct: a program for the graphical display of population structure. Mol Ecol Notes 4:137–138
Rousset F (1997) Genetic differentiation and estimation of gene flow from F-statistics under isolation by distance. Genetics 145:1219–1228
Simões PI, Lima AP, Magnusson WE (2008) Acoustic and morphological differentiation in the frog Allobates femoralis: relationships with the Upper Madeira River and other potential geological barriers. Biotropica 40:607–614
Sokal RR, Michener CD (1985) A statistical method for evaluating systematic relationships. Univ Kans Sci Bull 38:1409–1438
Stuart SN, Chanson JS, Cox NA, Young BE, Rodrigues ASL, Fischman DL, Waller RW (2004) Status and trends of amphibian declines and extinctions worldwide. Science 306:1783–1786
Tajima F (1989) Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. Genetics 123:585–595
Takezaki N, Nei M, Tamura K (2014) POPTREEW: web version of POPTREE for constructing population trees from allele frequency data and computing some other quantities. Mol Biol Evol 31:1622–1624
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729
Templeton AR, Crandall KA, Sing CF (1992) Cladistic analysis of phenotypic associations with haplotypes inferred from restriction endonuclease mapping and DNA sequence data. III. Cladogram estimation. Genetics 132:619–633
Tominaga A, Matsui M, Nakata K (2014) Genetic diversity and differentiation of the Ryukyu endemic frog Babina holsti as revealed by mitochondrial DNA. Zool Sci 31:64–70
Toyama M (1996) Rana (Babina) holsti Boulenger, 1982. In: Odate S (ed) Basic data for rare wild aquatic animals of Japan (III). Japan Fishers Resource Conservation Association, Tokyo
Van Oosterhout C, Hutchinson WF, Wills DPM, Shipley P (2004) Micro-Checker: software for identifying and correcting genotyping errors in microsatellite data. Mol Ecol Notes 4:535–538
Vences M, Andreone F, Glaw F, Kosuch J, Meyer A, Schaefer H-C, Veith M (2002) Exploring the potential of life-history key innovation: brook breeding in the radiation of the Malagasy treefrog genus Boophis. Mol Ecol 11:1453–1463
Vences M, Hauswald JS, Steinfartz S, Rupp O, Goesmann A, Künzel S et al (2013) Radically different phylogeographies and patterns of genetic variation in two European brown frogs, genus Rana. Mol Phylogenet Evol 68:657–670
Wake DB (2012) Facing extinction in real time. Science 335:1052–1053
Williamson-Natesan EG (2005) Comparison of methods for detecting bottlenecks from microsatellite loci. Conserv Genet 6:551–562
Wilson JP, Gallant JC (2000) Terrain analysis: principles and applications. Wiley, Hoboken
Wilson GA, Rannala B (2003) Bayesian inference of recent migration rates using multilocus genotypes. Genetics 163:1177–1191
Wright S (1943) Isolation by distance. Genetics 28:114–138
Wright S (1969) The theory of gene frequencies. University of Chicago Press, Chicago
Yokoyama Y, Lambeck K, De Deckker P, Johnson P, Fifield LK (2000) Timing of the last glacial maximum from observed sea-level minima. Nature 406:1998–2001
Acknowledgments
We are grateful to the Boards of Education of Okinawa Prefecture, Japan, for allowing us to collect the materials of Babina holsti protected by law. This work was supported by a Grants-in-Aid for Scientific Research (B and C; Nos. 24310173 and 20510216) to MS and a Grant-in-Aid for Young Scientists (B; No. 23710282) to TI from the Ministry of Education, Culture, Sports, Science and Technology, Japan.
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Kakehashi, R., Igawa, T. & Sumida, M. Genetic population structure and demographic history of an endangered frog, Babina holsti . Conserv Genet 16, 987–1000 (2015). https://doi.org/10.1007/s10592-015-0718-0
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DOI: https://doi.org/10.1007/s10592-015-0718-0


