Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Optimum Condition for Mass Culture of Hairy Roots from Artemisia sylvatica MAX

국내 자생 그늘쑥 (Artemisia sylvatica MAX) 모상근의 대량배양을 위한 최적조건

  • Shin, Sun-Hee (School of Life Science, College of Natural Science, Chungbuk National University) ;
  • Yang, Deok-Cho (School of Life Science, College of Natural Science, Chungbuk National University)
  • 신선희 (충북대학교 자연과학대학 생명과학부) ;
  • 양덕조 (충북대학교 자연과학대학 생명과학부)
  • Published : 2003.03.01
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Abstract

This research aims the production of anti-tumor substances through in vitro culture of hairy roots transformed by Agrobacterium rhizogenes in Artemisia sylvatica MAX and the effect of culture conditions on optimum growth of hairy roots. We investigated the optimum medium, pH, carbon source, sucrose, light, Fe and polyamine conditions of various lines of hairy roots (NK3, NK4, YX. NK3-10) induced from Artemisia sylvatica to increase the optimum growth of hairy roots. MS medium was the best for optimum growth of hairy root clone, NK3-S10. The optimum culture period was 4 weeks for NK3-S10. The optimum sucrose concentration was 3.5%. The optimum concentration of FeSO$_4$, spermine and spermidine was 0.1 mM, 10 mM and 100 mM, respectively.

본 실험은 그늘쑥을 Agrobacterium을 통해 형질전환체인 모상근을 유기하고 모상근의 대량배양을 위해 모상근이 가장 잘 자라는 생장 조건을 찾기 위해 수행되었다. 실험 결과를 종합해 보면 선발된 그늘쑥 모상근 세포주의 대량 생산을 위한 최적 배지는 MS 기본 배지임을 확인하였다. 또한 최적의 havesting time 모상근의 성장률이 좋고 갈변과 배지의 미생물 오염도가 적은 4주가 적당한 것으로 판단된다. 그리고 광 처리 하에서 모상근을 배양하는 것보다는 암상태에서 모상근을 키우고 pH는 5.8 정도가 탄소원은 sucrose로 3.5%가 모상근 을 대량배양시 최적조건임을 확인되었다. 미량원소인 Fe$^{2＋}$은 0.1 mM을 처리하는 것이 모상근의 성장을 촉진하였으며 spermine과 spermidine은 각각 10 mM와 100 mM로 처리하는 것이 모상근의 성장을 높일 수 있다. 앞으로 연구는 이 결과를 토대로 모상근 대량 배양을 위한 scale up조건과 bioreator의 설계에 관한 연구가 필요하다.

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References

  1. Avery MA, Jennings-White C, Chong WKM (1987) The total synthesis of (+)-artemisinin and (+)-9 dimethylartemisinin. Tetrahedron Lett 28: 4926-4632
  2. Choi CH (1998) Tropane alkaloids production in transgenic tissue induced by Agrobacterium. PhD thesis, Chungbuk National University, Cheongju, 92-100
  3. Choi CH, Kim YH,Yang DC (1999) Effect of media, culture periods, sucrose and dextrose on tropane alkaloid production in hairy root cultures of Hyoscyamus niger L. Korean J Plant Tiss Cult 26: 77-83
  4. Choi HY (1997) Physiological characteristics of genseng hairy root via light energy, MS thesis, Chungbuk National University, Cheongju, 55-61
  5. Cohen SS (1971) Introductiom to polyamines. Prentice-Hall, New Jersey 179
  6. Eu SL (1994) Establishment of cell and tissue culture systems and investigations of medicinal substances in Artemisia princeps var. MS thesis, Chungnam National University, Daejeon, 10-17
  7. Kang HM (1997) Tropane alkaloid production from transformed tissue of Datura Spp., MS thesis, Chungbuk National University, Cheongju, 58-62
  8. Kim YH, Yun KY, Yang DC (1998) Effect of media and elicitor on betalain and phytoaccsides production in hairy root cultures of Phytolacca esculenta van Houtte, Korean J Plant Tiss Cult 25: 507-514
  9. Lee HS, Kim KY, Kwon SY, Kwak SS (2002) Development of industrial transgenic plant using antioxidant enzyme genes, J Plant Biotech 29: 69-77 https://doi.org/10.5010/JPB.2002.29.2.069
  10. Lee SH, Kang HM, Song HC, Lee HS, Lee UC, Son KH, Kim SH, Kwon BM (2000) Sesquiterpene lactones, Inhibitors farnesyl protein transferase isolated from the flower of Artemisia syvatica, Terahedron 56: 4711-4715 https://doi.org/10.1016/S0040-4020(00)00394-X
  11. Liu CZ, Chen GZ, WangYC, Fan (2002) Effect of light irradiation on hairy root growth and artemisinin biosynthesis of Artemisia annuaL. Biochemistry 38: 581-585
  12. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tabacco tissue, Physiol Plant 15:473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  13. Norma B, Planiego, Giulietti M (1996) Aritemisinin production by Artemisia annua L.-transformed organ cultures, Enzyme Microb TechnoI 18: 526-530 https://doi.org/10.1016/0141-0229(95)00216-2
  14. Pack SW, Kim KH, Jung DY, Min TJ (1999) Cytotoxicity of Artemisia argyi extract against H9 (ATCC HTB 176) cell and antioxidant enzyme activities, Yankhak Hoeji 43: 598-60
  15. Palavan N, Gaslston AW (1982) Polyamine biosynthesis and titer during various development stage of Phaseolus vulgaris. Physiol Plant 55: 438-444 https://doi.org/10.1111/j.1399-3054.1982.tb04524.x
  16. Payne J, Hamill JD, Robins R, Rhodes MJC (1987) Production of hyoscyamine by hariy root cultures, Planta Med 53: 474-4785 https://doi.org/10.1055/s-2006-962776
  17. Schenk RU, Hildebrandt AC (1972) Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures, Can J Bot 50: 199-204 https://doi.org/10.1139/b72-026
  18. Song NH, Yang KJ, Bae CH, Yang DC (2001) Analysis and culture conditions for biosynthesis of polyacetylene from callus of ginsengsuperior lines, Korean J Plant Tiss Cult 28:123-128
  19. White PR (1943) A handbook of plant tissue culture, Jacques Cottell Press, Lancaster, Pennsylvania
  20. Yamada Y, Yun DJ, Hashimoto T (1994) Advances in plant biotechnology, Elsevier, 83-93
  21. Yang DC, Kim YH, Choi KM, Yun KY, Yang DC (1999) Effect of electron acceptor, electron transport inhibitors and antioxidants on growth and ginsenosides production in hary root cultures of Panax ginseng C.A. Meyer. J Ginseng Res 23: 205-210
  22. Xu XX, Zhu J, Huang DZ, Zhou WS (1986) Total synthesis of artemisinin and deoxiartemisinin. Tetrahedron Lett 42: 819-824 https://doi.org/10.1016/S0040-4020(01)87487-1