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Cryptic clonal lineages and genetic diversity in the loach Misgurnus anguillicaudatus (Teleostei: Cobitidae) inferred from nuclear and mitochondrial DNA analyses

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Abstract

In the loach Misgurnus anguillicaudatue, the asexual lineage, which produces unreduced clonal diploid eggs, has been identified. Among 833 specimens collected from 54 localities in Japan and two localities in China, 82 candidates of other lineage(s) of cryptic clones were screened by examining RFLP (restriction fragment length polymorphism)-PCR haplotypes in the control region of mtDNA. This analysis was performed because triploid loaches arise from the accidental incorporation of the sperm nucleus into unreduced diploid eggs of a clone. The categorization of members belonging to three newly identified lineages (clones 2–4) and the previously identified clonal lineage (clone 1) was verified by evaluating the genetic identity between two or more individuals from each clonal lineage based on RAPD (random amplified polymorphic DNA)-PCR and multilocus DNA fingerprints. We detected 75 haplotypes by observing the nucleotide status at variable sites from the control region of mtDNA. Phylogenic trees constructed from such sequences showed two highly diversified clades, A and B, that were beyond the level common for interspecific genetic differentiation. That result suggests that M. anguillicaudatus in Japan is not a single species entity. Two clone-specific mtDNA sequences were included in clade A, and the loaches with such sequences may be the maternal origin of the clones.

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References

  • Arai K (2001) Genetic improvement of aquaculture finfish species by chromosome manipulation techniques in Japan. Aquaculture 197:205–228

    Article  CAS  Google Scholar 

  • Arai K (2003) Genetics of the loach, Misgurnus anguillicaudatus: recent progress and perspective. Folia Biol (Krakow) 51(Suppl):107–117

    Google Scholar 

  • Arai K, Matsubara K, Suzuki R (1991a) Karyotype and erythrocyte size of spontaneous tetraploidy and triploidy in the loach Misgurnus anguillicaudatus. Nippon Suisan Gakkaishi 57:2157–2172

    Google Scholar 

  • Arai K, Matsubara K, Suzuki R (1991b) Chromosomes and developmental potential of progeny of spontaneous tetraploid loach Misgurnus anguillicaudatus. Nippon Suisan Gakkaishi 57:2173–2178

    Google Scholar 

  • Arai K, Matsubara K, Suzuki R (1993) Production of polyploids and viable gynogens using spontaneously occurring loach, Misgurnus anguillicaudatus. Aquaculture 117:227–235

    Article  Google Scholar 

  • Arai K, Mukaino M (1997) Clonal nature of gynogenetically induced progeny of triploid (diploid × tetraploid) loach Misgurnus anguillicaudatus (Pisces: Cobitididae). J Exp Zool 278:412–421

    Article  CAS  Google Scholar 

  • Arias-Rodriguez L, Morishima K, Arai K (2007) Genetically diversified population in the loach Misgurnus anguillicaudatus, inferred from newly developed microsatellite markers. Mol Ecol Notes 7:82–85

    Article  CAS  Google Scholar 

  • Asahida T, Kobayashi T, Saitoh K, Nakayama I (1996) Tissue preservation and total DNA extraction from fish stored at ambient temperature using buffer containing high concentration of urea. Fish Sci 62:727–730

    Google Scholar 

  • Beukeboom LW, Vrijenhoek RC (1998) Evolutionary genetics and ecology of sperm-dependent parthenogenesis. J Evol Biol 11:755–782

    Article  Google Scholar 

  • Boroń A (2000) Cytogenetic characterization of the loaches of the genera Sabanejewia, Misgurnus and Barbatula (Pisces, Cobitidae). Folia Zool 49(Suppl 1):37–44

    Google Scholar 

  • Boroń A (2003) Karyotypes and cytogenetic diversity of the genus Cobitis (Pisces, Cobitidae) in Poland: a review. Cytogenetic evidence for a hybrid origin of some Cobitis triploids. Folia Biol (Krakow) 51(Suppl):49–54

    Google Scholar 

  • Dawley RM (1989) An introduction to unisexual vertebrates. In: Dawley RM, Bogart HP (eds) Evolution and ecology of unisexual vertebrates. New York State Museum, Bull. 466, New York, pp 1–18

    Google Scholar 

  • Dawley RM, Goddard KA (1988) Diploid-triploid mosaics among unisexual hybrids of the minnows Phoxinus eos and Phoxinus neogaeus (Pisces:Cyprinidae). Evolution 42:649–659

    Article  Google Scholar 

  • Dawley RM, Schultz RJ, Goddard KA (1987) Clonal reproduction and polyploidy in unisexual hybrids of Phoxinus eos and Phoxinus neogaeus (Pisces; Cyprinidae). Copeia 1987:275–283

    Article  Google Scholar 

  • Echelle AA, Echelle AF, Crozier CD (1983) Evolution of an all-female fish, Menidia clarkhubbsi (Atherinidae). Evolution 37:772–784

    Article  Google Scholar 

  • Echelle AA, Mosier DT (1982) Menidia clarkhubbsi, n. sp. (Pisces: Atherinidae) an all-female species. Copeia 1982:533–540

    Article  Google Scholar 

  • Fersenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–781

    Article  Google Scholar 

  • Goddard KA, Dawley RM (1990) Clonal inheritance of a diploid nuclear genome by a hybrid freshwater minnow (Phoxinus eos-neogaeus, Pisces: Cyprinidae). Evolution 44:1052–1065

    Article  Google Scholar 

  • Goddard KA, Megwinoff I, Wessner LL, Giaimo F (1998) Confirmation of gynogenesis in Phoxinus eos-neogaeus (Pisces; Cyprinidae). J Hered 89:151–157

    Article  Google Scholar 

  • Goddard KA, Schultz RJ (1993) Aclonal reproduction by polyploid members of the clonal hybrid species Phoxinus eos-neogaeus (Cyprinidae). Copeia 1993:650–660

    Article  Google Scholar 

  • Itono M, Morishima K, Fujimoto T, Bando E, Yamaha E, Arai K (2006) Premeiotic endomitosis produces diploid eggs in the natural clone loach, Misgurnus anguillicaudatus (Teleostei: Cobitidae). J Exp Zool 305A:513–523

    Article  CAS  Google Scholar 

  • Itono M, Okabayashi N, Morishima K, Fujimoto T, Yoshikawa H, Yamaha E, Arai K (2007) Cytological mechanisms of gynogenesis and sperm incorporation in unreduced diploid eggs of the clonal loach, Misgurnus anguillicaudatus (Teleostei:Cobitidae). J Exp Zool 307A:35–50

    Article  CAS  Google Scholar 

  • Janko K, Kotlik P, Rab P (2003) Evolutionary history of asexual hybrid loaches (Cobitis: Teleostei) inferred from phylogenetic analysis of mitochondrial DNA variation. J Evol Biol 16:1280–1287

    Article  PubMed  CAS  Google Scholar 

  • Janko K, Culling MA, Rab P, Kotlik P (2005) Ice age cloning – comparison of the Quaternary evolutionary histories of sexual and clonal forms of spiny loaches (Cobitis; Teleostei) using the analysis of mitochondrial DNA variation. Mol Ecol 14:2991–3004

    Article  PubMed  CAS  Google Scholar 

  • Khan MR, Arai K (2000) Allozyme variation and genetic differentiation in the loach, Misgurnus anguillicaudatus. Fish Sci 66:211–222

    Article  CAS  Google Scholar 

  • Kitagawa T, Watanabe M, Kitagawa E, Yoshioka M, Kashiwagi M, Okazaki T (2003) Phylogeography and the maternal origin of the tetraploid form of the Japanese spined loach, Cobitis biwae, revealed by mitochondrial DNA analysis. Ichthyol Res 50:318–325

    Article  Google Scholar 

  • Kitagawa T, Jeon S-R, Kitagawa E, Yoshioka M, Kashiwagi M, Okazaki T (2005) Genetic relationships among the Japanese and Korean striated spined loach complex (Cobitidae: Cobitis) and their phylogenetic positions. Ichthyol Res 52:111–122

    Article  Google Scholar 

  • Kumar S, Tamura K, Nei M (2004) MEGA4: integrated software for molecular evolutionary genetic analysis and sequence alignment. Brief Bioinform 5:150–163

    Article  PubMed  CAS  Google Scholar 

  • Lusk S, Kosco J, Halacka K, Luskova V, Flajshans M (2003) Identification of Cobitis from the Slovakian part of the Tisza basin. Folia Biol (Krakow) 51(Suppl):61–65

    Google Scholar 

  • Monaco PJ, Rasch EM, Balsano JS (1984) Apomictic reproduction in the Amazon molly, Poecilia formosa, and its triploid hybrids. In: Dawley RM, Bogart HP (eds) Evolution and ecology of unisexual vertebrates. New York State Museum, Bull. 466, New York, pp 311–328

    Google Scholar 

  • Morishima K, Nakayama I, Arai K (2001) Microsatellite-centromere mapping in the loach, Misgurnus anguillicaudatus. Genetica 111:59–69

    Article  PubMed  CAS  Google Scholar 

  • Morishima K, Horie S, Yamaha E, Arai K (2002) A cryptic clonal line of the loach Misgurnus anguillicaudatus (Teleostei:Cobitidae) evidenced by induced gynogenesis, interspecific hybridization, microsatellite genotyping and multilocus DNA fingerprinting. Zool Sci 19:565–575

    Article  PubMed  Google Scholar 

  • Morishima K, Oshima K, Horie S, Fujimoto T, Yamaha E, Arai K (2004) Clonal diploid sperm of the diploid-triploid mosaic loach, Misgurnus anguillicaudatus (Teleostei:Cobitidae). J Exp Zool 301A:502–511

    Article  Google Scholar 

  • Oliva O, Hensel K (1961) Some remarks on eastern Asiatic loaches of the genus Misgurnus (Cobitidae). Jpn J Ichthyol 8:86–91

    Google Scholar 

  • Palumbi SR, Martin A, Remano S, McMillan WO, Stice L, Grabowski G (1991) Simple fool’s guide to PCR, ver 2.0. Zoology Department, University of Hawaii, Honolulu, USA

    Google Scholar 

  • Posada D, Crandall KA (2001) Selecting the best fit model of nucleotide substitution. Syst Biol 50:580–601

    Article  PubMed  CAS  Google Scholar 

  • Quatro JM, Avise JC, Vrijenhoek RC (1992a) An ancient clonal lineage in fish genus Poeciliopsis (Atheriniformes: Poeciliidae). Proc Nat Acad Sci USA 89:348–352

    Article  Google Scholar 

  • Quatro JM, Avise JC, Vrijenhoek RC (1992b) Mode of origin and sources of genotypic diversity in triploid gynogenetic fish clones (Poeciliopsis, Poeciliidae). Genetics 130:621–628

    Google Scholar 

  • Rab P, Rabova M, Bohren J, Lusk S (2000) Genetic differentiation of the two hybrid diploid-polyploid complexes of loaches, genus Cobitis (Cobitidae) involving C. taenia, C. elongatoides and C. spp. in the Czech Republic: karyotypes and cytogenetic diversity. Folia Zool 49(Suppl):55–66

    Google Scholar 

  • Saitoh K (1989) Asian pond loach. In: Kawanabe H, Mizuno N (eds) Freshwater fishes of Japan. Yamakei Pub, Tokyo, pp 382–385

    Google Scholar 

  • Sakai T, Mihara M, Shitara S, Yonekawa H, Hosoya K, Miyazaki J (2003) Phylogenetic relationships and intraspecific variations of loaches of the genus Lefua (Balitoridae, Cypriniformes). Zool Sci 20:501–514

    Article  PubMed  CAS  Google Scholar 

  • Sakaizumi M, Shimizu Y, Matsuzaki T, Hamaguchi S (1993) Unreduced diploid eggs produced by interspecific hybrids between Oryzias latipes and O. curvinotus. J Exp Zool 266:312–318

    Article  Google Scholar 

  • Shimizu Y, Shibata N, Sakaizumi M, Yamashita M (2000) Production of diploid eggs through premeiotic endomitosis in the hybrid medaka between Oryzias latipes and O. curvinotus. Zool Sci 17:951–958

    Article  Google Scholar 

  • Slechtova V, Luskova V, Slechta V, Lusk S, Halacka K, Bohlen J (2000) Genetic differentiation of the two hybrid diploid-polyploid complexes of loaches, genus Cobitis (Cobitidae), involving C. taenia, C. elongatoides and C. spp. in the Czech Republic: allozyme interpopulation and interspecific differences. Folia Zool 49(Suppl):67–78

    Google Scholar 

  • Tamura K, Nei M (1993) Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol Biol Evol 10:512–526

    PubMed  CAS  Google Scholar 

  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins HG (1997) The Clustal X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 24:4876–4882

    Article  Google Scholar 

  • Vasil’ev VP, Vasil’eva ED, Oshinov AG (1989) Evolution of diploid-triploid-tetraploid complex in fishes of the genus Cobitis. In: Dawley RM, Bogart HP (eds) Evolution and ecology of unisexual vertebrates. New York State Museum, Bull. 466, New York, pp 153–169

    Google Scholar 

  • Vrijenhoek RC (1989) Genetic and evolutionary constraints on the origin and establishment of unisexual vertebrates. In: Dawley RM, Bogart HP (eds) Evolution and ecology of unisexual vertebrates. New York State Museum, Bull. 466, New York, pp 24–31

    Google Scholar 

  • Vrijenhoek RC, Dawley RM, Cole CJ, Bogart HP (1989) A list of the known unisexual vertebrates. In: Dawley RM, Bogart HP (eds) Evolution and ecology of unisexual vertebrates. New York State Museum, Bull. 466, New York, pp 19–23

    Google Scholar 

  • Welsh J, McClelland M (1990) Fingerprinting genomes using PCR with PCR with arbitrary primers. Nucleic Acid Res 18:7213–7218

    Article  PubMed  CAS  Google Scholar 

  • Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acid Res 18:6531–6533

    Article  PubMed  CAS  Google Scholar 

  • Zhang Q, Arai K (1999) Distribution and reproxductive capacity of natural triploid individuals and occurrence of unreduced eggs as a cause of polyploidization in the loach, Misgurnus anguillicaudatus. Ichthyol Res 46:153–161

    Article  Google Scholar 

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Acknowledgments

This study was partly supported by Grants-in-Aid for the 21st Century COE (Center of Excellence) Program from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan and for the Scientific Research (B) (No.18380108) from the Japanese Society for the Promotion of Science (JSPS) to KA. A part of this study was also supported by the JSPS Ronpaku (Dissertation Ph.D.) Program (CSC-10610) to YL and the JSPS Invitation Fellowship for Research in Japan (S-05135) to AB.

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  1. Kagayaki Morishima

    Present address: Fisheries Laboratory, Kinki University, 153 Shirahama, Wakayama, 649-2211, Japan

Authors and Affiliations

  1. Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido, 041-8611, Japan

    Kagayaki Morishima, Yuka Nakamura-Shiokawa, Etsuko Bando & Katsutoshi Arai

  2. Department of Life Science and Technology, Dalian Fisheries University, Dalian, 116023, China

    Ya-Juan Li

  3. Department of Zoology, University of Warmia and Mazury in Olsztyn, Oczapowski 5 St., Olsztyn, 10-718, Poland

    Alicja Boroń

  4. Department of Fisheries Biology and Genetics, Bangladesh Agriculture University, Mymensingh, 2202, Bangladesh

    Md. Mukhlesur Rahman Khan

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  1. Kagayaki Morishima
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Correspondence to Kagayaki Morishima.

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Morishima, K., Nakamura-Shiokawa, Y., Bando, E. et al. Cryptic clonal lineages and genetic diversity in the loach Misgurnus anguillicaudatus (Teleostei: Cobitidae) inferred from nuclear and mitochondrial DNA analyses. Genetica 132, 159–171 (2008). https://doi.org/10.1007/s10709-007-9158-1

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