Effect of NO3- and NH4+ Concentrations on Root Growth and Eleutherosides Accumulation in adventitious root Culture of Eleutherococcus senticosus

가시오갈피의 부정근 배양시 부정근의 생육과 eleutheroside류의 함량에 미치는 NO3-와 NH4+ 비율 및 농도의 영향

  • Ahn, Jin-Kwon (Department of Forest Genetic Resources, Korea Forest Research Institute) ;
  • Lee, Wi-Young (Department of Forest Genetic Resources, Korea Forest Research Institute) ;
  • Park, Young-Ki (Department of Forest Genetic Resources, Korea Forest Research Institute)
  • 안진권 (국립산림과학원 산림유전자원부) ;
  • 이위영 (국립산림과학원 산림유전자원부) ;
  • 박영기 (국립산림과학원 산림유전자원부)
  • Received : 2006.11.03
  • Accepted : 2007.03.07
  • Published : 2007.03.31

Abstract

This study was carried out to investigate the effect of $NO_3{^-}$ and $NH_4{^+}$ on the adventitious root growth and eleuthroside synthesis of Eleutherococcus senticosus during 3 L-bioreactor culture. The change of medium component ratio was also measured during culture. The fresh weignt of adventitious root reached to the greatest level of 24.4g FW/L in the presence of 50 mM $NO_3{^-}$ and 10 mM $NH_4{^+}$, representing 3.4-fold increase compared to the 60 mM $NH_4{^+}$. However, as the increase of the portion of $NH_4{^+}$, the root growth was decreased. Maximum eleutheroside B and E1 production were $249{\mu}g/g$ and $43{\mu}g/g$, respectively, with 30 mM total nitrogen source. The maximum production of eleutheroside E were $788{\mu}g/g$ with 120 mM total nitrogen source. The greatest weight of adventitious root increased up to 6.2 fold of inoculum size within 9 weeks. The change of pH was influenced from 4.81 to 6.35 and the amounts of $NH_4{^+}$ and $K^+$ were decreased during culture periods. From these results we suggest, need further study of various treatment to increase the growth of biomass and the accumulation of useful secondary metabilites.

생물반응기를 이용하여 가시오갈피 부정근 증식시 부정근의 생장과 유용 이차대사산물인 eleuthroside 류 생산에 미치는 $NO_3{^-}$$NH_4{^+}$의 영향 및 배양기간에 따른 배지성분의 변화량을 분석하였다. 부정근 증식은 질소농도가 50 mM $NO_3{^-}$와 10 mM $NH_4{^+}$ 농도비율로 참가된 처리구에서 최대(24.4g FW/L)에 이르러 60 mM $NH_4{^+}$ 첨가에 비해서는 3.4배 증식되었다. 또한 $NH_4{^+}$의 첨가농도비율이 높아질수록 부정근의 증식은 감소하였다. Eleutheroside B와 E1은 총 질소량이 30 mM 처리에서 각각 $249{\mu}g/g$$43{\mu}g/g$을 생산하여 가장 우수하였으나, eleutheroside E는 총 질소농도가 높아질수록 생산량도 높아져 120 mM에서 $788{\mu}g/g$로 가장 우수하였다. 부정근 생장은 초기 접종량 대비 배양 9주만에 6.2배가 증식되었고, 배양기간별 배지 성분의 변화에서는 산도는 4.81-6.35로 변화가 비교적 심했고, $NH_4{^+}$$K^+$는 배양기간이 경과할수록 함유량이 낮았다. 이러한 결과로 보아 부정근의 생장과 유용이차대사산물의 함량을 높이기 위하여 다양한 처리의 연구가 수행되어야 할 것으로 생각된다.

Keywords

References

  1. Ahn, J.K., Park, S.Y., Lee, W.Y., and Park, Y.P. 2006. Effect of jasmonic acid on root growth and eleutheroside accumulation in adventitious root culture of Eleutherococcus koreanum. Journal of Korean Forest Society. 95(1): 32-37
  2. Ahn, J.K., Park, S.Y., Lee, W.Y., and Lee, J.J. 2005. Effect of growth regulators on adventitious root growth and eleutherosides and chlorogenic acid accumulation in air lift bioreactor culture of Eleutherococcus koreanum. Korean Journal of plant biotechnology. 32(1): 57-61 https://doi.org/10.5010/JPB.2005.32.1.057
  3. Ahn, J.K., Lee, W.Y., and Park, S.Y. 2003. Effect nitrogen source on the cell growth and production of secondary metabolites in bioreactor cultures of Eleutherococcus senticosus. Korean Journal of Plant Biotechnology. 30(3): 301-305 https://doi.org/10.5010/JPB.2003.30.3.301
  4. Ahn, J.K., Lee, W.Y., Oh, S.J., and Park, Y.H. 2000. The contents of chlorogenic acid and eleutheroside E in Eleutherococcus senticosus (Rupr. et Maxim.) Harms. Journal of Korean Forest Society. 89: 216-222
  5. Akalezi, C.O., Liu, S., Li, Q.S., Yu, J.T., and Zhong, J.J. 1999. Combined effects of initial sucrose concentration and inoculum size on cell growth and ginseng saponin production by suspension cultures of Panax ginseng. Process Biochemistry. 34: 639-642 https://doi.org/10.1016/S0032-9592(98)00132-0
  6. Fujita, Y., Hara, Y., Ogino, T., and Suga, C. 1981. Production of shikonin dervatives by cell suspension cultures of Lithospermum erythrorhizon. 1. Effect of nitrogen sources on the production of shikonin dervatives. Plant Cell Reports. 1: 59-60 https://doi.org/10.1007/BF00269272
  7. Gorret, N., Rosli, S.K., Oppenheim, S.P., Willis, L.B., Lessard, P.A., Rha, C.K., and Sinskey, A.J. 2004. Bioreactor culture of oil palm (Elaeis guineensis) and effects of nitrogen source, inoculum size, and conditioned medium on biomass production. Journal of Biotechnology. 108: 253-263 https://doi.org/10.1016/j.jbiotec.2003.12.009
  8. Han, J.Y., and Choi, Y.E. 2003. Mass production of Eleutherococcus senticosus plants through in vitro cell culture. Korean Journal of Plant Biotechnology. 30: 167-172 https://doi.org/10.5010/JPB.2003.30.2.167
  9. Kang, S.M., Jung, H.Y., Kang, Y.M., Yun, D.J., Bahk, J.D., Yang, J.K., and Choi, M.S. 2004. Effects of methyl jasmonate and salicylic acid on the production of tropane alkaloids and the expression of PMT and H6H in adventitious root cultures of Scopolia parviflora. Plant Science. 166: 745-751 https://doi.org/10.1016/j.plantsci.2003.11.022
  10. Kang, J.S., Linh, P.T., Cai, X.F., Kim, H.S., Lee, J.J., and Kim, Y.H. 2001. Quantitative determination of eleutheroside Band E from Acanthopanax species by high performance liquid chromatography. Archives pharmacal research. 24: 407-411 https://doi.org/10.1007/BF02975184
  11. Liu, S., and Zhong, J.J. 1997. Simultaneous production of ginseng saponin and polysaccharide by suspension cultures of Panax ginseng: Nitrogen effects. Enzyme and Microbial. Technology. 21: 518-524
  12. Liu, S., and Zhong. J.J. 1998. Phosphate effect onproduction of ginseng saponin and polysaccharide by cell sus-pension cultures of Panax ginseng and Panax quinquefolium, Process Biochemistry. 33: 69-74 https://doi.org/10.1016/S0032-9592(97)00064-2
  13. Murashige, T., and Skoog. F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum. 15: 473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  14. Panda, A.K., Mishra, S., and Bisaria, V.S. 1992. Alkaloid production by plant cell suspension cultures of Holarrhena antidysenterica : 1. Effect of major nutrients. Biotechnology Bioeng. 39: 1043-1051 https://doi.org/10.1002/bit.260391008
  15. Sakuta, M., Takagi, T., and Komamine, A. 1987. Effects of nitrogen source on betacyanin accumulation and growth in suspension cultures of Phytolacca americana. Physiology Plant. 71: 459-463 https://doi.org/10.1111/j.1399-3054.1987.tb02884.x
  16. Slacanin, I., Marston, A., and Hostettmann, K. 1991. The isolation of Eleutherococcus senticosus constituents by centrifugal partition chromatography and their quantitative determination by high performance liquid chromatography. Phytochemistry Analysis. 2: 137-142 https://doi.org/10.1002/pca.2800020310
  17. Son, S.H., Choi, S.M., Choi, K.B., Lee, Y.H., Lee, D.S., Choi, M.S., and Park, Y.G. 1999. Selection and proliferation of rapid growing cell lines from embryo derived cell cultures of Yew tree (Taxux cuspidata Sieb. et Zucc). Biotechnology Bioprocess Engineering. 4: 112-118 https://doi.org/10.1007/BF02932380
  18. Son, S.H., Choi, S.M., Lee, Y.H., Choi, K.B., Yun, S.R., Kim, J.K., Park, H.J., Kwon, O.W., Noh, E.W., Seon, J.H., and Paek, K.Y. 2000. Large-scale growth and taxane production in cell cultures of Taxus cuspidata (Japanese yew) using a novel bioreactor. Plant Cell Reports. 19: 628-633 https://doi.org/10.1007/s002990050784
  19. Suresh, B., Thirnmaraju, R., Bhagyalakshmi, N., and Ravishankar, G.A. 2004. Polyamine and methyl jasmonateinfluenced enhancement of betalaine production in hairy root cultures of Beta vulgaris grown in a bubble column reactor and studies on efflux of pigments. Process Biochemistry. 39: 2091-2096 https://doi.org/10.1016/j.procbio.2003.10.009
  20. Yu, K.W., Gao, W., Hahn, E.J., and Paek, K.Y. 2002. Jasmonic acid improves ginsenoside accumulation in adventitious root culture of Panax ginseng C.A. Meyer. Biochemical Engineering Journal. 11: 211-215 https://doi.org/10.1016/S1369-703X(02)00029-3