Geographic Variation in Pond Smelt (Hypomesus nipponensis) by RAPD Analysis

RAPD 분석에 의한 빙어 (Hypomesus nipponensis)의 지리적 변이

  • Kim, Yong-Ho (Faculty of Marine Life Science, College of Ocean Science and Technology, Kunsan National University) ;
  • Park, Su-Young (Department of Aquatic Life Medicine, College of Ocean Science and Technology, Kunsan National University) ;
  • Yoon, Jong-Man (Department of Aquatic Life Medicine, College of Ocean Science and Technology, Kunsan National University)
  • 김용호 (국립군산대학교 해양과학대학 해양생명과학부) ;
  • 박수영 (국립군산대학교 해양과학대학 수산생명의학과) ;
  • 윤종만 (국립군산대학교 해양과학대학 수산생명의학과)
  • Received : 2005.10.20
  • Accepted : 2006.02.24
  • Published : 2006.03.31

Abstract

Genomic DNA isolated from two geographical populations of pond-smelt (Hypomesus nipponensis) was amplified for RAPD (randomly amplified polymorphic DNA) analysis. The populations were obtained from Chungju (CJ), in the inland area, and Dangjin (DJ), in the vicinity of the West Sea in Korea. Seven arbitrarily selected primers, OPB-06, OPB-10, OPB-13, OPB-17, OPC-09, OPC-17 and OPC-20, were used to generate the shared loci, polymorphic, and specific loci. Three hundred and eighty-three loci observed per primer were identified in the CJ population, and 287 were identified in the DJ population. Among them, 91 polymorphic loci or 23.8% were polymorphic in the CJ population, and 47 (16.4%) in the DJ population. The number of shared loci observed was 198 in the CJ population and 176 in the DJ population. Forty-four and 75 specific loci were detected in the CJ and DJ populations, respectively. Especially, 99 numbers of shared loci by the two populations, with an average of 14.1 per primer, were observed in the two pond-smelt populations. The average bandsharing value between the two geographical pond-smelt populations was $0.700{\pm}0.008$, ranging from 0.600 to 0.846. Compared separately, the bandsharing value of individuals within the CJ population was higher than that of the DJ population. The dendrogram obtained using the data from the seven primers indicated three genetic clusters: cluster 1, CJ 01, 02, 03, 04, 05, 06, 07, 08, 09, 10, and 11; cluster 2, DJ 01, 02, 03, 04, 05, 06, 07, 08, and 09; and cluster 3, DJ 10 and 11. The genetic distance between the two geographical populations ranged from 0.040 to 0.545. Thus, RAPD-PCR analysis revealed a significant genetic distance between the two pond-smelt populations.

RAPD 분석을 하기 위해서 우리나라 내륙의 충주지역과 서해에 인접한 당진지역에서 빙어 (Hypomesus nipponensis)의 두 지리적 집단으로부터 genomic DNA 를 분리 추출하였다. OPB-06, OPB-10, OPB-13, OPB-17, OPC-09, OPC-17 및 OPC-20의 7개 primer를 사용하여 shared loci, polymorphic 및 specific loci를 확인하였다. 충주 빙어집단에서는 한 primer 당 383개의 loci가 관찰되었고, 당진 집단에서는 287개의 loci가 확인되었다. 관찰된 loci 중에 23.8%에 해당되는 91개의 polymorphic loci가 충주 집단에서 확인되었고, 당진 집단에서는 47 (16.4%)개가 확인되었다. 각 집단에서 공유하는 loci의 수는 각각 충주 빙어집단에서 198개 그리고 당진 집단에서는 176개로 관찰되었다. 충주 빙어집단과 당진 집단에서는 각각 44개와 75개의 specific loci가 나타났다. 특히 두 집단이 공유하는 loci의 수는 99개로서 primer 당 평균 14.1개로 확인되었다. 두 빙어 집단의 bandsharing value의 평균값은 $0.700{\pm}0.008$로서 0.600에서 0.846의 범위를 나타내었다. 각각을 비교해 보면, 충주 집단에 속한 개체의 bandsharing value의 평균값이 당진 집단에서의 값보다 높게 나타났다. 7개의 primer를 사용하여 얻어진 dendrogram은 cluster 1 (CJ 01, 02, 03, 04, 05, 06, 07, 08, 09, 10 및 11), cluster 2 (DJ 01, 02, 03, 04, 05, 06, 07, 08 및 09) 및 cluster 3 (DJ 10 및 11)와 같이 3개의 유전적 클러스터로 나뉘어졌다. 두 집단의 유전적 거리는 0.040에서 0.545 사이로 나타났다. 따라서 RAPD-PCR 분석을 통해서 빙어 두 집단의 유의성이 있는 유전적 거리를 확인하였다.

Keywords

Acknowledgement

Supported by : Kunsan National University

References

  1. Cagigas, M.E., E. Vazquez, G. Blanco and J.A. Sanchez. 1999. Combined assessment of genetic variability in populations of brown trout (Salmo trutta L.) based on allozymes, microsatellites, and RAPD markers. Mar. Biotechnol., 1 : 286-296 https://doi.org/10.1007/PL00011778
  2. Callejas, C. and M.D. Ochando. 1998. Identification of Spanish barbel species using the RAPD technique. J. Fish Biol., 53 : 208-215 https://doi.org/10.1111/j.1095-8649.1998.tb00121.x
  3. Chenyambuga, S.W., O. Hanotte, J. Hirbo, P.C. Watts, S.J. Kemp, G.C. Kifaro, P.S. Gwakisa, P.H. Petersen and J.E.O. Rege. 2004. Genetic characterization of indigenous goats of sub-Saharan Africa using microsatellite DNA markers. Asian-Aust. J. Anim. Sci., 17 : 445-452 https://doi.org/10.5713/ajas.2004.445
  4. Huang, B.X., R. Peakall and P.J. Hanna. 2000. Analysis of genetic structure of blacklip abalone (Haliotis rubra) populations using RAPD, minisatellite and microsatellite markers. Mar. Biol., 136 : 207-216 https://doi.org/10.1007/s002270050678
  5. Iyengar, A., S. Piyapattanakorn, D.M. Dtone, D.A. Heipel, B.R. Howell, S.M. Baynes and N. Maclean. 2000. Identification of microsatellite repeats in turbot (Scophthalmus maximus) and dover sole (Solea solea) using a RAPD-based technique: Characterization of microsatellite markers in dover sole. Mar. Biotechnol., 2 : 49-56
  6. Jeffreys, A.J. and D.B. Morton. 1987. DNA fingerprints of dogs and cats. Anim. Genet., 18 : 1-15 https://doi.org/10.1111/j.1365-2052.1987.tb00739.x
  7. Kim, I.S., Y. Choi, C.L. Lee, Y.J. Lee, B.J. Kim and J.H. Kim. 2005. Illustrated Book of Korean Fishes. Kyo-Hak Pub., Seoul, 150 pp
  8. Kim, J.Y., C.Y. Park and J.M. Yoon. 2004. Genetic differences and DNA polymorphism in oyster (Crassostrea spp.) analysed by RAPD-PCR. Korean J. Genet., 26 : 123-134
  9. Klinbunga, S., A. Boonyapakdee and B. Pratoomchat. 2000. Genetic diversity and species-diagnostic markers of mud crabs (Genus Scylla) in Eastern Thailand determined by RAPD analysis. Mar. Biotechnol., 2 : 180- 187
  10. Koh, M.C., C.H. Lim, S.B. Chua, S.T. Chew and S.T. Phang. 1998. Random amplified polymorphic DNA (RAPD) fingerprints for identification of red meat animal species. Meat Sci., 48 : 275-285 https://doi.org/10.1016/S0309-1740(97)00104-6
  11. Liu, Z., P. Li, B.J. Argue and R.A. Dunham. 1998. Inheritance of RAPD markers in channel catfish (Ictalurus punctatus), blue catfish (I. furcatus) and their $F_{1}$, $F_{2}$ and backcross hybrids. Anim. Genet., 29 : 58-62 https://doi.org/10.1046/j.1365-2052.1998.00284.x
  12. Mamuris, Z., C. Stamatis, M. Bani and C. Triantaphyllidis. 1999. Taxonomic relationships between four species of the Mullidae family revealed by three genetic methods: allozymes, random amplified polymorphic DNA and mitochondrial DNA. J. Fish Biol., 55 : 572-587 https://doi.org/10.1111/j.1095-8649.1999.tb00700.x
  13. McCormack, G.C., R. Powell and B. Keegan. 2000. Comparative analysis of two populations of the brittle star Amphiura filiformis (Echinodermata: Ophiuroidae) with different life history strategies using RAPD markers. Mar. Biotechnol., 2 : 100-106
  14. Park, C.Y. and J.M. Yoon. 2005. Genetic differences and variation in two largehead hairtail (Trichiurus lepturus) populations determined by RAPD-PCR analysis. Korean J. Ichthyol., 17 : 173-186
  15. Partis, L. and R.J. Wells. 1996. Identification of fish species using random amplified polymorphic DNA (RAPD). Mol. Cell. Probes, 10 : 435-441 https://doi.org/10.1006/mcpr.1996.0060
  16. Smith, P.J., P.G. Benson and S.M. McVeagh. 1997. A comparison of three genetic methods used for stock discrimination of orange roughy, Hoplostethus atlanticus: allozymes, mitochondrial DNA, and random amplified polymorphic DNA. Fish. Bull., 95 : 800-811
  17. Tassanakajon, A., S. Pongsomboon, P. Jarayabhand, S. Klinbunga and V. Boonsaeng. 1998. Genetic structure in wild populations of black tiger shrimp (Penaeus monodon) using randomly amplified polymorphic DNA analysis. J. Mar. Biotechnol., 6 : 249-254
  18. Waldbieser, G.C. and W.R. Wolters. 1999. Application of polymorphic microsatellite loci in a channel catfish Ictalurus punctatus breeding program. J. World Aquacult. Soc., 30 : 256-262 https://doi.org/10.1111/j.1749-7345.1999.tb00873.x
  19. Welsh, J. and M. McClelland. 1990. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res., 18 : 7213-7218 https://doi.org/10.1093/nar/18.24.7213
  20. Welsh, J., C. Petersen and M. McClelland. 1991. Polymorphisms generated by arbitrarily primed PCR in the mouse: application to strain identification and genetic mapping. Nucleic Acids Res., 19 : 303-306 https://doi.org/10.1093/nar/19.2.303
  21. Yoon, J.M. 2001. Genetic similarity and difference between common carp and Israeli carp (Cyprinus carpio) based on random amplified polymorphic DNAs analyses. Korean J. Biol. Sci., 5 : 333-339 https://doi.org/10.1080/12265071.2001.9647624
  22. Yoon, J.M. and G.W. Kim. 2001. Randomly amplified polymorphic DNA-polymerase chain reaction analysis of two different populations of cultured Korean catfish Silurus asotus. J. Biosci., 26 : 641-647 https://doi.org/10.1007/BF02704762
  23. Yoon, J.M. and H.Y. Park. 2002. Genetic similarity and variation in the cultured and wild crucian carp (Carassius carassius) estimated with random amplified polymorphic DNA. Asian-Aust. J. Anim. Sci., 15 : 470-476 https://doi.org/10.5713/ajas.2002.470
  24. Yoon, J.M. and Y.H. Kim. 2003a. Wide marsh clam (Corbicula spp.) populations from three sites analysed by RAPD-PCR-AGE. Bull. Electrochem., 19 : 337-348
  25. Yoon, J.M. and G.W. Kim. 2003b. Genetic differences between cultured and wild penaeid shrimp (Penaeus chinensis) populations analysed by RAPD-PCR. Korean J. Genet., 25 : 21-32
  26. Yoon, J.M. and J.Y. Kim. 2004. Genetic differences within and between populations of Korean catfish (S. asotus) and bullhead (P. fulvidraco) analysed by RAPD-PCR. Asian-Aust. J. Anim. Sci., 17 : 1053-1061 https://doi.org/10.5713/ajas.2004.1053