생명과학회지 (Journal of Life Science)

  • 제22권11호
  • /
  • Pages.1470-1476
  • /
  • 2012
  • /
  • 1225-9918(pISSN)
  • /
  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
권호 표지
DOI QR코드

DOI QR Code

RAPD와 ITS 영역에 의한 민자주방망이 버섯의 유전적 변이

Genetic Variation Based on Random Amplified Polymorphic DNA (RAPD) and Internal Transcribed Spacer (ITS) Region Sequences in Lepista nuda

  • 이양숙 (대구한의대학교 한약자원학과) ;
  • 김남우 (대구한의대학교 한약자원학과) ;
  • 김종봉 (대구가톨릭대학교 의생명과학과)
  • Lee, Yang Suk (Department of Herbal Biotechnology, Daegu Haany University) ;
  • Kim, Nam Woo (Department of Herbal Biotechnology, Daegu Haany University) ;
  • Kim, Jong Bong (Department of Medical Life Science, Catholic University of Daegu)
  • 투고 : 2012.08.02
  • 심사 : 2012.11.23
  • 발행 : 2012.11.30
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

본 연구는 유럽에서 식용버섯으로 선호도가 높은 Lepista nuda (민자주방망이버섯)에 대하여 random amplified polymorphic DNA (RAPD)와 internal transcribed spacer (ITS) 염기서열을 이용하여 종내 및 종간의 유전적 변이를 분석하였다. RAPD 분석 결과 40개의 random primer 중 다형성을 나타내는 primer는 22개 였으며, 증폭된 밴드는 355개, DNA 단편의 크기는 200~4,000 bp의 사이에 위치하였다. RAPD band들을 marker로 하여 Nei-Li's의 방법을 이용한 비유사도 지수행렬을 조사한 결과 L. nuda 종내 유전적 변이는 0~21.60%로 나타났으며, L. nuda와 L. sordida의 종간에는 16.93~24.82%, L. irina와는 20.62~25.54%로 나타났으며, L. sordida와 L. irina와의 종간 변이는 23.49%로 나타났다. ITS I 과 II 영역의 673 bp의 염기서열을 분석하여 비유사도 지수행렬을 조사한 결과, L. nuda의 종내 변이는 1.58~11.47%였으며, L. nuda와 L. sordida와는 3.83~12.88%로 나타났다. 그리고 L. nuda와 L. irina는 7.11~15.61%였으며, L. sordida와 L. irina와의 종간 변이는 4.79%로 나타났다. 본 실험결과 RAPD와 ITS실험을 통해 확인된 primer와 연기서열은 Lepista속의 종을 검색 및 분류 시 유전적 표지 marker로서 이용 할 수 있을 것으로 생각된다.

A genetic variation of Lepista nuda and two genus Lepista species (L. irina and L. sordida) were analyzed by random amplified polymorphic DNA (RAPD) and internal transcribed spacer (ITS) sequence analysis. In the resulting RAPD analysis, 22 out of 40 random primers amplified polymorphic RAPD fragment patterns, the amplified bands were 355, and DNA fragment sizes were 200-400bp. Intraspecific genetic dissimilarity of the 10 L. nuda strains were calculated to range from 0% to 21.60%, L. sordida from 16.93% to 24.82%, L. irina were 20.62% to 25.54%, and intraspecific genetic dissimilarity of L. sordida and L. irina was 23.49%. The 673 base pairs were sequenced during the analysis of the ITS I and II region; six L. nuda strains intraspecific genetic dissimilarities ranged from 1.58% to 11.47%, L. nuda and L. sordida from 3.83% to 12.88%, L. nuda and L. irina from 7.11% to 15.61%, and intra-specific genetic variation between L. sordida and L. irina was 4.79%. The findings showed that RAPD and ITS sequencing could be used for developing molecular genetic markers and screening of unidentified genus Lepista species.

키워드

참고문헌

  1. Alam, N., Shim, M. J., Lee, M. W., Shin, P. G., Yoo, Y. B. and Lee, T. S. 2009. Physiologenetic relationship in different commercial strains of Pleurotus nebredonsis based on ITS sequence and RAPD. Mycobiology 37, 183-188. https://doi.org/10.4489/MYCO.2009.37.3.183
  2. Cheong, J. C., Lee, M. C., Kim, B. G., Park, D. S., Hong, S. B. and Park, J. S. 2004. Interspecific distinguish ability of veiled lady mushrooms (Dictyophora spp.) based on rDNA-ITS analysis. Kor. J. Mycol. 32, 1-7. https://doi.org/10.4489/KJM.2004.32.1.001
  3. Dulge, B., Ergul, C. C. and Gucin, F. 2002. Antimicrobial activity of the macrofungus Lepista nuda. Fitoterapa 3, 695-697.
  4. Kim, J. B., and Hwang, S. G. 2001. A study on the mycelia culture and genetic characteristics in Lepista nuda. Korean J. Life Sci. 1, 496-501.
  5. Kim, J. H. 2005. Purification and characterization of fibrinolytic enzyme from Lepista nuda. Kor. J. Mycol. 33, 69-74. https://doi.org/10.4489/KJM.2005.33.2.069
  6. Kim, J. H., Yoo, K. H., Kim, Y. S., Seok, S. J. and Kim, Y. S. 1998. The screening of fibrinolytic activities of extracts from mushrooms in Mt. Chiak. Kor. J. Mycology 26, 589-593.
  7. Kim, S H., and Kim, J. B. 2004. Genetic variation of Korean Lepista nuda. J. Life Sci. 14, 115-120. https://doi.org/10.5352/JLS.2004.14.1.115
  8. Lee, S. B., Milgroom, M. G. and Taylor, J. W. 1988. A rapid, high yield mini-prep method for isolation of total genomic DNA from fungi. Fungal Genetics Newsletter 35, 23-24.
  9. Lee, S. S., Choi, K. J. and Oh, C. H. 1996. Sawdust cultures of Lepista nuda. Kor. J. Mycol. 24, 274-279.
  10. Lee, Y. S., Park, D. C., Joo, E. Y., Shin, S. R. and Kim, N. W. 2005. Study on the antioxidant activity of the extracts from the Lepista nuda. J. Korean Soc. Food Sci. Nutr. 34, 942-917. https://doi.org/10.3746/jkfn.2005.34.7.942
  11. Lee, Y. S., Han, J. Y., Joo, E. Y., Shin, S. R. and Kim, N. W. 2005. Study on the antitumor effects of extracts from Lepista nuda Mushroom. J. Korean Soc. Food. Nutr. 34, 317-322. https://doi.org/10.3746/jkfn.2005.34.3.317
  12. Lim, Y. J., Lee, C. Y., Park, J. E., Kim, S. W., Lee, H. S. and Ro, H. S. 2010. Molecular genetic classification of Hypsizigus marmoreus and development of strain-specific DNA markers. Kor. J. Mycol. 38, 34-39. https://doi.org/10.4489/KJM.2010.38.1.034
  13. Moradali, M. F., Mostafavi, H., Ghods, S. and Hedgaroude, G. A. 2007. Immunomodulating and anticancer agent int the realm of macromycetes fungi (macrofungi). Int. Immunopharmacol. 7, 701-724. https://doi.org/10.1016/j.intimp.2007.01.008
  14. Nei, M. 1973. Genetic distance between populations. America Nature 106, 283-292.
  15. Norkrans, B. 1950. Studies in growth and cellulolytic enzymes of Tricholoma. Bot. Ups. 11, 1-126.
  16. Park, D. S., Kang, H. W., Kim, K. T., Cho, S. M., Park, Y. J., Shin, H. S., Lee, M. B. and Go, S. J. 2001. PCR-based sensitive detection of wood decaying fungus Phellinus linteus by specific primer from rDNA ITS regions. Micobiology 29, 7-10.
  17. Park, H. G., Ko, H. G., Kim, H. S. and Park, M. W. 2004. Molecular identification of Asian isolates of medicinal mushroom Hericium erinaceum by phylogenetic analysis of nuclear ITS rDNA. J. Microbiol. Biotechnol. 14, 816-821.
  18. Park, W. H. 1998. Studies on inorganic components of Korean wild edible mushrooms trace mineral elements of Armillariella mella, Hygrophorous russula, Armillariella tabescens, Lepista nuda, Lepista sordida, Hygrocybe conica. Kor. J. Mycol. 21, 273-278.
  19. Saitou, N. and Nei, M. 1987. The neighbour-joining method: A new method for reconstruction phylogenetic trees. Mol. Biol. Evol. 4, 406-425.
  20. Smith, L. M. Sanders, J. Z., Kaiser, R. J., Hughers, P., Dodd, C., Conner, C. R., Heiner, C., Kent, S. B. H. and Hood, L. E. 1986. Fluorescence detection in automated DNA sequence analysis. Nature 321. 674-679. https://doi.org/10.1038/321674a0
  21. Sneath, R. R. and Sokal, R. R. 1973. Numerical taxonomy. Freeman, San Francisco. pp. 573.
  22. Stott, K., Desmerger, C. and Holford, P. 2005. Relationship among Lepista species determined by CAPS and RAPD. Mycol. Res. 109, 205-211. https://doi.org/10.1017/S0953756204001662
  23. Stott, K. and Broderick, A. 1996. Response of Australian strains of the mushroom L. nuda to temperature and substrate. J. Janic, Progress in New Crops. In ASHS press Arlington. VA. pp. 476-479.
  24. Wasser, S. P. 2002. Medical mushrooms as a source of antitumor and immunomodulating polysaccharides. Appl. Microbiol. Biotechnol. 60, 258-274. https://doi.org/10.1007/s00253-002-1076-7
  25. Waugh, R. and Powell, W. 1992. Using RAPD markers for crop improvement. TIBTECH. 10, 186-190. https://doi.org/10.1016/0167-7799(92)90212-E
  26. White, T. J., Bruns, T., Lee, S. and Taylor, J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR protocols-A guide to methods and application. Eds., Innis, M. A., Gelfand, D. H., Sninsky, J. J. and White, T. J. Academic Press Inc., San diego, California. pp. 315-322.
  27. Williams, J. G., Kubelik, A. R., Livak, K. J., Rafalski, J. A. and Tingey, S. V. 1990. DNA polymorphism amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18, 6531-6535. https://doi.org/10.1093/nar/18.22.6531

피인용 문헌

  1. Secondary Structure of the Ribosomal Internal Transcribed Spacer (ITS) Region of Hypsizygus marmoreus vol.23, pp.10, 2013, https://doi.org/10.5352/JLS.2013.23.10.1260