Genetic Identification of Hybrids between Rhodeus uyekii and R. notatus by Sequence Analysis of RAG-1 Gene

RAG-1 유전자의 염기서열 분석에 의한 각시붕어 Rhodeus uyekii와 떡납줄갱이 R. notatus 잡종의 동정

  • Yun, Young-Eun (Department of Marine Biotechnology, Soonchunhyang University) ;
  • Lee, Il-Ro (Department of Marine Biotechnology, Soonchunhyang University) ;
  • Park, Sang-Yong (Department of Marine Biotechnology, Soonchunhyang University) ;
  • Kang, Eon-Jong (Southern Regional Inland Research Institute, NFRDI) ;
  • Kim, Eung-O (Southern Regional Inland Research Institute, NFRDI) ;
  • Yang, Sang-Keun (Southern Regional Inland Research Institute, NFRDI) ;
  • Nam, Yoon-Kwon (Department of Aquaculture, Pukyong National University) ;
  • Bang, In-Chul (Department of Marine Biotechnology, Soonchunhyang University)
  • 윤영은 (순천향대학교 해양생명공학과) ;
  • 이일로 (순천향대학교 해양생명공학과) ;
  • 박상용 (순천향대학교 해양생명공학과) ;
  • 강언종 (국립수산과학원 남부내수면연구소) ;
  • 김응오 (국립수산과학원 남부내수면연구소) ;
  • 양상근 (국립수산과학원 남부내수면연구소) ;
  • 남윤권 (부경대학교 양식학과) ;
  • 방인철 (순천향대학교 해양생명공학과)
  • Published : 2009.02.25


Reciprocal interspecific hybrids between two bitterling species Rhodeus uyekii (RU) and R. notatus (RN) were genetically identified based on the partial sequence analysis of recombination activating gene-1 (RAG-1) gene. Out of 863 bp positions analyzed, 13 nucleotide substitutions were detected between the two parental species (RU and RN genotypes). Both the induced hybrids (RU female$\times$RN male; UN genotype) and their reciprocal counterparts (RN female$\times$RU male; NU genotype) displayed the double peaks (or polymorphism) of sequence chromatograms at the 13 diagnostic positions, indicating that those hybrids were actual karyogamy derived from the two parental haploid genomes. However, it was not possible to distinguish between the reciprocal interspecific hybrids.

한국 고유종인 각시붕어 R. uyekii와 떡납줄갱이 R. notatus로부터 유도된 정교배 및 상반교배 잡종어류의 분자생물학적 동정을 위하여 핵에서 encoding되는 RAG-1 유전자의 염기서열 분석을 실시하였다. 분석된 863 bp의 염기서열 중 각시붕어와 떡납줄갱이 사이에는 총 13개의 위치에서 염기서열 변이가 탐색되었다. 잡종어류의 RAG-1 유전자 염기서열을 분석한 결과 모계와 부계의 염기서열 차이를 보인 13개의 변이 부분에서 부모의 염기서열을 다같이 반영하는 double peaks 패턴을 보였으나 정교매체(UN 유전형)와 상반교배체(NU 유전형) 간의 염기서열 차이는 관찰되지 않았다.


  1. Asahida, T., T. Kobayashi, K. Saitoh and I. Nakayama, 1996. Tissue preservation and total DNA extraction from fish stored at ambient temperature using buffers containing high concentration of urea. Fish. Sci., 62, 727-730
  2. Busse, K., J. Freyhof and A. Zarske, 1999. Serum-electrophoretic pattems of various cyprinid hybrids from the rivers Elbe and Sieg (Teleostei: Cyprinidae). Abh. Mus. Tierk. Dresden., 21, 265-272
  3. Candolin, U. and J. D. Reynolds, 2002. Why do males tolerate sneakers? Tests with the European bitterling, Rhodezιs serzceus. Behav. Ecol. Sociobiol., 51, 146-152
  4. Cebrat, M., A. Cebula, A. Laszk:iewicz,M. Kasztura, A. Miazek and P. Kisielow, 2008. Mechanism of Iymphocyte-specific inactivation of RAG-2 intragenic promoter of NWC: Implications for epigenetic control of RAG locus. Mol. Immunol., 45, 2297-2306
  5. Groth, J. G and G F. Barrowclough, 1999. Basal divergences in birds and the phylogenetic utility of the nuclear RAG-I gene. Mol. Phyl. Evol., 12, 115-123
  6. Hall, T. A., 1999. BioEdit: A user-friendly biogical sequence aligment editor and analysis program for windows 95/98/NT. Nucl. Acids. Symp. Ser., 41, 95-98
  7. Jansson, H., I. Holmgren, K. Wedin and T Andersson, 1991. High frequency of natural hybrids between Atlantic salmon, Salmo salar L., and brown trout, S. trutta L., in a Swedish river. J. Fish Biol., 39, 343-348
  8. Kang, E.-J., C.-H. Kim, I.-S. Park, H. Yang and Y.-C. Cho, 2006. Early developmental charaterristics of Induced hybrids between Rhodeus uyekii and R. notaus (Pisces: Cyprinidae). Kor. J. Ichthyol., 18, 339-346. (in Korean)
  9. Kang, E.-J., E.-M. Kim, Y. J. Kim, S. G Lee, D. S. Sim, Y.-H. Kim and I.-S. Park, 2005. Effect of lidocaine hydrochloride and clove oil as anaesthetic on Korean rose bitterling, Rhodeus uyekii and bitterling, Acheilognathus koreensis. J. Aquacult., 18, 272-279. (in Korean)
  10. Kanoh, Y., 2000. Reproductive success associated with territoriality, sneaking, and grouping in male rose bitterlings, Rhodeus ocellatus (Pisces: Cyprinidae). Environ. Biol. Fishes., 57, 143- 154
  11. Kawamura, K. and K. Hosoya, 2000. Masculinization mechanism ofhybrids in bitterlings (Teleostei: Cyprinidae). J. Hered., 91, 464-473
  12. Kim, I.-J., 1997. A osteological study of Rhodeus uyekii. Kor. J. Ichthyol., 9, 130-140. (in Korean)
  13. Kim, H. S., Y. H. Kim, J.-Y. Jo, G. H. Yoon and B.-S. Ha, 1999a. Nuptial color component of Korean rose bitterling (Rhodeus uryekii). J. Kor. Fish. Soc., 32, 520-524. (in Korean)
  14. Kim, H. S., Y. H. Kim, S. H. Cho and J.-Y. Jo, 1999b. Effects of dietary carotenoids on the nuptial color of the bitterling (Rhodeus uyekii). J. Kor. Fish. Soc., 32, 276-279. (in Korean)
  15. Kim, Y. U. and K. H. Han, 1990. Early life history of the Korean bitterling, Rhodeus uyekii (Cyprinidae), with a note on minute tubercles on the skin surface. Kor. J. Ichthyol., 2, 159-168. (in Korean)
  16. Kuriiwa, K., N. Hanzawa, T. Yoshino, S. Kimura and M. Nishida, 2007. Phylogenetic relationships and natural hybridization in rabbitfishes (Teleostei: Siganidae) inferred from mitochondrial and nuclear DNA analyses. MoL Phyl. Evol., 45, 69-80
  17. Quenouille, B., E. Bermingham and S. Planes 2004. Molecular systematic of the damselfishes (Teleostei: Pomacentridae): Bayesian phylogenetic analyses of mitochondrial and nuclear DNA sequences. Mol. Phyl. Evol., 31, 66-88
  18. Slechtova, V., J. Bohlen and H.H. Tan, 2007. Families of Cobitoidea (Teleostei: Cypriniformes) as revealed from nuclear genetic data and the position of the mysterious genera Barbucca, Psilorhynchus, Swepenticobitis and Vaillantella. Mol. Phyl. Evol., 44, 1358-1365
  19. Sonnenberg, R., A. Nolte and D. Tautz, 2007. An evaluation of LSU rDNA DI-D2 sequences for their use in species identification. Front. Zool., 4, 6
  20. Suzuki, N. and S.-R. Jeon, 1988a. Hybridization experiments in acheilognathine fishes (Cyprinidae), an intrageneric hybrid between Rhodeus uryekii and R. ocellatus smithi. Kor. J. Limnol., 21, 129-142. (in Korean)
  21. Suzuki, N. and S.-R. Jeon, 1988b. Hybridization experiments in acheilognathine fishes (Cyprinidae), an intrageneric hybrid between Rhodeus suigensis and R. uyekii. Kor. J. Limnol., 21, 57-78. (in Korean)
  22. Suzuki, N., N. Akiyama and T. Hibiya, 1985. Development ofthe bitterling, Rhodeus uyekii (Cyprinidae), with a note on minute tubercles on the skin surface. Jap. J. Ichthyol., 32, 28-34
  23. Thompson, J. D., T. J. Gibson, F. Plewniak, F. Jeanmougin and D. G. Higgins, 1997. The CLUSTAL X windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res., 25, 4876-4882
  24. Wendel, J. F. and J. J. Doyle, 1998. Phylogenetic incongruence: Window into genome history and molecular evolution. In: Soltis, D.E., Soltis, P.S., Doyle, J.J. (Eds.), Molecular Systematics of Plants II: DNA Sequencing. Kluwer Academic Publishers, Boston, pp. 265-296