Korean Journal of Environment and Ecology (한국환경생태학회지)

Korean Society of Environment and Ecology (한국환경생태학회)
권호 표지

A Study of Potential of Diet Analysis in the Korean Water Deer(Hydropotes inermis argyropus) using Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis(PCR-DGGE)

고라니의 식이물 분석에 있어 Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis(PCR-DGGE)의 이용 가능성 연구

  • Park, Ji-Eun (Division of EcoScience, Ewha Womans University) ;
  • Kim, Baek-Jun (Conservation Genome Resource Bank for Korean Wildlife(CGRB), College of Veterinary Medicine and BK21 Program for Veterinary Science, Seoul National University) ;
  • Lee, Sang-Don (Department of Environmental Science and Engineering, College of Engineering, Ewha Womans University)
  • 박지은 (이화여자대학교 에코과학부) ;
  • 김백준 (서울대학교 수의과대학 수의학과) ;
  • 이상돈 (이화여자대학교 공과대학 환경공학과)
  • Received : 2009.10.30
  • Accepted : 2010.06.25
  • Published : 2010.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

The aim of this study is to examine feeding habits of the Korean water deer(Hydropotes inermis argyropus) from its rumen contents using a PCR-DGGE method. For this study, rumen contents were collected from water deer causalities by natural death or road-kill in two different sites(Cheorwon, Gangwon province and the Eastern part of Jeonnam province). DNA was extracted from rumen contents of a total of 44 individuals. Two primers, rbcLZ1aF(GC) and rbcL19bR, were used for PCR amplifications of ribulose-1,5-bisphosphate carboxylase large subunit (rbcL) gene. Among 44 samples, twenty-nine samples were successfully amplified by PCRs. The 29 PCR products of partial rbcL gene were applied for PCR-DGGE. Totally, six families of plants were detected from the diet analyses. Five families of plants were found in Cheorwon, Gangwon province, but only three families of plants were found in the Eastern part of Jeonnam province. The PCR-DGGE method will provide us with a potential tool to study feeding habits of ungulates including water deer, even though our results failed to identify the prey plants at the level of species.

이 연구의 목적은 고라니(Hydropotes inermis argyropus)의 위내용물을 대상으로 PCR-DGGE 방법을 이용하여 그 식이습성을 조사하는데 있다. 이 연구를 위해, 강원도 철원과 전라남도 동부지역 등에서 자연사 혹은 로드킬에 의해 죽은 고라니 사체의 위에서 식이물 샘플을 채취하였다. 총 44개체의 위내용물에서 각각 DNA를 추출하였고, 두 가지의 프라이머(rbcLZ1과 rbcL19bR)를 이용하여 ribulose-1,5-bisphosphate carboxylase large subunit(rbcL) gene을 PCR 증폭하였다. 44개의 샘플 중 29 샘플에서 성공적으로 PCR을 수행하였다. 이 29개 partial rbcL gene의 PCR product는 PCR-DGGE에 이용되었다. 식이물에 대한 분석결과 총 6과의 식물이 확인되었다. 강원도 철원의 경우, 5과가 나타난 반면, 전라남도 동부의 경우, 3과만이 확인되었다. 이 연구에서는 종수준의 먹이식물의 구별에는 실패하였 지만, 차후 이 PCR-DGGE 기법은 고라니를 포함한 초식동물의 식이습성을 분석하는데 하나의 가능성 있는 방법이 될 것으로 생각된다.

Keywords

References

  1. Butzler, W.(l990) Grzimek's Encyclopedia Mammals, Volume five. McGraw-Hill Publishing Company, New York, pp. 198-199.
  2. Chamrad, A.D. and T.W. Box(1964) A point frame for sampling rumen contents. J. Wildl. Manage. 28: 473-477. https://doi.org/10.2307/3798199
  3. Cooke, A. and L. Farrell(1998) Chinese Water Deer. The Mammal Society, London and the British Deer Society, Fordingbridge, pp. 1-32.
  4. Deagle, B.E., S.N. Jarman, D. Pemberton and N.J. Gales(2005a) Genetic screening for prey in the gut contents from a Giant squid. J. Hered. 96: 417-423. https://doi.org/10.1093/jhered/esi036
  5. Deagle, B.E., D.J. Tollit, S.N. Jarman, M.A. Hindell, A.W. Trites and N.J. Gales(2005b) Molecular scatology as a tool to study diet: analysis of prey DNA in scats from captive Steller sea lions. Mol. Ecol. 14: 1831-1842. https://doi.org/10.1111/j.1365-294X.2005.02531.x
  6. Degarine-wichatitsky, M., Y. Soubeyran, D. Maillard and P. Duncan(2005) The diets of introduced rusa deer(Cervus timorensis russa) in a native sclerophyll forest and a native rainforest of New Caledonia. New Zealand J. Zool. 32: 117-126. https://doi.org/10.1080/03014223.2005.9518403
  7. Hanley, T.A., D.E. Spalinger, K.A. Hanley and J.W. Schoen(1985) Relationships between fecal and rumen analyses for deer diet assessments in Southeastern Alaska. Northwest Science 59: 10-16.
  8. Hofreiter, M., H.N. Poinar, W.G. Spaulding, K. Bauer, P.S. Martin, G. Possnert and S. Paabo(2000) A molecular analysis of ground sloth diet through the last glaciation. Mol. Ecol. 9: 1975-1984. https://doi.org/10.1046/j.1365-294X.2000.01106.x
  9. Kim, E.K.(2007) Food diet analysis of Korean water deer using a microhistological method. M.S. thesis, Kangwon Univ., Rep. of Korea. pp. 1-49.
  10. Lee, B.K.(2003) Morphological, Ecological and DNA Taxonomic Characteristics of Chinese Water Deer(Hydropotes inermis Swinhoe). Ph.D. thesis, Chungbuk national Univ., Rep. of Korea, pp. 1-101.
  11. Lee, J.H., I.D. Lee and H.S. Lee(1990) Studies on the Utilization of Browse by the Sika Deer(Cervus nippon). Korean J. Anim. Sci. 32: 100-108.
  12. Muyzer, G., E.C. de Waal and A.G. Uitterlinden(1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction amplified gene coding for 16S rRNA. Applied and Environ. Microbiol. 59: 695-700.
  13. Ramirez, R.G., J.B. Quintanilla and J. Aranda(1997) White-tailed food habits in northeastern Mexico. Small Rum. Res. 25: 141-146. https://doi.org/10.1016/S0921-4488(96)00960-1
  14. Seiki, T. and U. Hironori(2007) Meso-scale variation in winter food composition of sika deer in Tochigi Prefecture, central Japan. Mammal Study 32: 115-120. https://doi.org/10.3106/1348-6160(2007)32[115:MVIWFC]2.0.CO;2
  15. Seo, M.H.(2006) Analysis of Asiatic Black Bear's foods by using feces. M.S. thesis, Kookmin Univ., Rep. of Korea, pp. 1-53.
  16. Tollit, D.J., A.D. Schulze, A.W. Trites, P.F. Olesiuk, S.J. Crockford, T.S. Gelatt, R.R. Ream and K.M. Miller(2009) Development and application of DNA techniques for validating and improving pinniped diet estimates. Ecol. Appl. 19: 889-905. https://doi.org/10.1890/07-1701.1
  17. Wemmer, C.(1998) Deer, status survey and conservation action plan. IUCN, SSC deer specialist group, 69pp.
  18. Won, C.M. and K.G. Smith(1999) History and current status of mammals of the Korean peninsula. Mam. Rev. 29: 3-36. https://doi.org/10.1046/j.1365-2907.1999.00034.x
  19. Yang, B.G.(2002) Systematics, ecology and current population status of the goral, Naemorhedus caudatus, in Korea. Ph.D. thesis, Chungbuk national Univ., Rep. of Korea. pp. 1-137.
  20. Zhang, E.(1998) Uniparental female care in the Chinese water deer at Whipsnade Wild animal park, England. Acta Theriol. Sinica 18: 178-183.