Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Effects of Plant Growth Regulators, Medium Salt Strength and Nitrogen Ratio on Cell Culture of Gymmema sylvestre

식물생장조절물질, 무기물 농도 및 질소원 비율이 Gymmma sylvestre 세포 배양에 미치는 영향

  • Lee, Eun-Jung (Research Center for the Development of Advanced Horticultural Technology, Chungbuk National University) ;
  • Han, Eun-Joo (Research Center for the Development of Advanced Horticultural Technology, Chungbuk National University) ;
  • Paek, Kee-Yoeup (Research Center for the Development of Advanced Horticultural Technology, Chungbuk National University)
  • 이은정 (충북대학교 첨단원예기술개발연구센터) ;
  • 한은주 (충북대학교 첨단원예기술개발연구센터) ;
  • 백기엽 (충북대학교 첨단원예기술개발연구센터)
  • Published : 2006.06.30
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Abstract

This study was carried out to investigate the effects of plant growth regulators, medium salt strength and nitrogen ratio on cell culture of Gymnema sylvestre. Cell growth was inhibited by 2,4-D higher than 1.0 mg L$^{-1}$, but not by kinetin lower than 0.5 mg L$^{-1}$. Maximal cell growth was obtained at 1.0 mg L$^{-1}$ 2,4-D and 0.1 mg L$^{-1}$ kinetin. Cell growth was greatest at 1x MS medium but high strength of MS medium inhibited cell growth due to low water potential in the medium. In $NH_4^+:NO_3^-$ ratio of 0:60 (i.e. 0.0 mM $^NH_4^+$ and 60.0 mM $NO_3^-$), cells growth was highest but cells were smaller and whiter compared with those in other $NH_4^+:NO_3^-$ ratio. Reduced cell growth was observed with continuous culture. These results suggested that optimal cell culture of G. sylvestre could be achieved with 1x MS medium with 20:40 ratio of $NH_4^+:NO_3^-$ supplemented with 1.0 mg L$^{-1}$ 2,4-D and 0.1 mg L$^{-1}$ kinetin.

배지 내 식물생장조절물질, 무기물 농도, 질소원 비율이 G. sylvestre 세포배양에 미치는 영향을 알아보고자 실험을 수행하였다 1.0 mg L$^{-1}$ 이상2,4-D 처리시 세포 생장량이 급격히 감소한 반면, 0.5 mg L$^{-1}$ 이하 kinetin 처리구들은 세포 생장량이 큰 차이가 없었다. 따라서 G. sylvestre 세포배양의 경우 2,4-D 1.0 mg L$^{-1}$와 kinetin 0.1 mg L$^{-1}$를 함께 사용하는 것이 세포 생장량 증대에 효율적이라고 판단되었다. 무기물 농도가 G. sylvestre 세포 생장량에 미치는 영향을 조사한 결과 1x MS에서 세포 생장량이 가장 높았으며, 배지 내 무기물 농도가 1x MS 이상일 경우 배지의 낮은 수분포텐셜로 인해 세포생장이 크게 억제되었다. $NH_4^+:NO_3^-$ 비율이 0:60에서 가장높은 생장량을 나타내었으나 세포의 크기가 작고 백색을 나타내었으며 연속배양시 생장량이 점차 감소하므로, G. sylvestre 세포를 연속 배양할 경우 $NH_4^+:NO_3^-$를 20:40으로 사용하는 것이 효과적이라고 생각되었다.

Keywords

References

  1. Ahn JK, Lee WY, Park SY (2003) Effect of nitrogen source on the cell growth and production of secondary metabolites in bioreactor cultures of Eleutherococcus senticosus. Kor J Plant Biotechnol 30: 301-305 https://doi.org/10.5010/JPB.2003.30.3.301
  2. Ananthan R, Baskar C, NarmathaBai V, Pari L, Latha M, RamKumar KM (2003) Antidiabetic effect of Gymnema montanum leaves: effect on lipid peroxidation induced oxidative stress in experimental diabetes. Pharm Res 48: 551-556 https://doi.org/10.1016/S1043-6618(03)00219-6
  3. Ashok Kumar HG, Murthy HN, Paek KY (2002) Somatic embryogenesis and plant regeneration in Gymnema sylvestre. Plant Cell Tiss Org Cult 71: 85-88 https://doi.org/10.1023/A:1016529322046
  4. Attard EG, Scicluna-Spiteri A (2001) Ecballium elaterium an in vitro source of cucurbitacins. Fitoterap 72: 46-53 https://doi.org/10.1016/S0367-326X(00)00256-2
  5. Behrend J, Mateles IR (1975) Nitrogen metabolism in plant cell suspension cultures. Plant physiol 56: 584-589 https://doi.org/10.1104/pp.56.5.584
  6. Bryne TG, Hasek RF (1979) Poinettia leaf injury associated with $NH_4^+$ fertilization and low soil pH. Flor Rev 165: 65-68
  7. Chanwitheesuk A, Teerawutgulrag A, Rakariyatham N (2005) Screening of antioxidant activity and antioxidant compounds of some edible plants of Thailand. Food Chem 92: 491-497 https://doi.org/10.1016/j.foodchem.2004.07.035
  8. Choudhury BP (1988) Assessment and conservation of medicinal plants of hubaneswar and its neighbourhood. Indigenous Medicinal Plants, Today and Tomorrow's Printers and Publishers, New Delhi, pp 211-219
  9. Fonseka HD, Asanuma KI, Ichii M (1997) Changes in nitrate reductase activity of leaf and nitrogen distribution with growth in potato plants. Jap J Crop sci 65: 669-674
  10. Garcia G, Tagliaferro P, Ferri A, Evangelista AM, Duffard R, Brusco A (2004) Study of tyrosine hydroxylase immunoreactive neurons in neonate rats lactationally exposed to 2,4-dichlorophenoxyacetic acid. Neurotoxicol 25: 951-957 https://doi.org/10.1016/j.neuro.2004.05.004
  11. Gopi C, Vatsala TM (2006) In vitro studies on effects of plant growth regulators on callus and suspension culture biomass yield from Gymnema sylvestre R. Br. Afr J Biotechnol 5: 1215-1219
  12. Gueon DY, Park YI, Jun SS, Jin CD, Hong YN (2001) Plant physiology. In: Hopkins WG, (eds), Introduction to plant physiology, Eulyoo publishing company Ltd, Seoul, pp 69-86
  13. Hdider CH, Vezina LP, Ahn CH (1994) Short-term studies of $NO_3\;^-$ and $NH_4\;^+$ uptake by micropropagated strawberry shoots cultured with or without $CO_2$ enrichment. Plant Cell Tiss Org Cult 37: 185-191 https://doi.org/10.1007/BF00043614
  14. Khosroushahi AY, Valizadeh M, Ghasempour A, Khosrowshahli M, Naghdibadi H, Dadpour MR, Omile Y (2006) Improved Taxol production by combination of inducing factors in suspension cell culture of Taxus baccata. Cell Biology International 30: 262-269 https://doi.org/10.1016/j.cellbi.2005.11.004
  15. Kim GW, Kim CG, Park YG, Soo WY, Soon SH, Soon JG, Shim GB, Ahn YH, Eun JS, Lee VB, Lee JS, Lee CH, Lim HT, Jung JD, Ji SY, Hahn BH, Hahn EJ, Hang P (2003) Plant tissue culture. In: Paek KY, (eds), Plant tissue culture - technology, Hyang moon sa, Seoul, pp 59-63
  16. Kim YS (2002) Production of ginsenosides through bioreactor cultures of adventitious roots in ginseng (Panas ginseng C.A. Meyer). PhD thesis, Chungbuk National University, Cheongju
  17. Komalavalli N, Rao MN (2000) In vitro micropropagation of Gymnema sylvestre - A multipurpose medicinal plant. Plant Cell Tiss Org Cult 61: 97-105 https://doi.org/10.1023/A:1006421228598
  18. Lee EJ (2005) Effect of chemical and physical environment on cell culture of Gymnema sylvestre. MS thesis, Chungbuk National University, Cheongju
  19. Leidi EO, Silberbush M, Lips SH (1991) Wheat growth as affected by nitrogen type, pH and salinity. I. Biomass production and mineral composition. J Plant Nut 14: 235-246 https://doi.org/10.1080/01904169109364198
  20. Lim JN (2000) Methods of plant analysis. In: Lim JN, (eds), Methods of soil and plant analysis, National institute of agricultural science and technology, Seoul, pp 139-140
  21. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  22. Narayan MS, Thimmaraju R, Bhagyalakshmi N (2005) Interplay of growth regulators during solid-state and liquid-state batch cultivation of antocyanin cell line of Daucus carota. Proc Biochem 40: 351-358
  23. Park CH, Seong NS, Paek KY, Lee CH (1998) Micropropagation through callus culture in chinese foxglove (Rehmannia glutinosa). Kor J Plant Tiss Cult 25: 171-175
  24. Rokem JS, Goldberg J (1985) Secondary metabolites from plant cell suspension cultures: methods for yield improvement. In: Mizrahi A, Van Wezel AL, (eds), Advances in Biotechnological Processes, Ed 4, Alan R. Liss. Inc., New York, pp 241-274
  25. SAS Institute (1989) SAS / STAT User's Guide, Ed 4, Vol. 6, SAS Inst., Cary, NC
  26. Satdive RK, Abhilash P, Fulzele DP (2003) Antimicrobial activity of Gymnema sylvestre leaf extract. Fitoterapia 74: 699-701 https://doi.org/10.1016/S0367-326X(03)00154-0
  27. Selvanayagam ZE, Gnanavendhan SG, Chandrasekhran P, Balakrishna K, Rao RB (1995) Plants with antisnake venom activity - a review on pharmacological and clinical studies. Fitoterapia 65: 99-111
  28. Taguchi G, Yoshizawa K, Kodaira R, Hayashida N, Okazaki M (2001) Plant hormone regulation on scopoletin metabolism from culture medium into tobacco cells. Plant Sci 160: 905-911 https://doi.org/10.1016/S0168-9452(00)00464-7
  29. Wakiuchi N, Matsumoto H, Takahashi E (1971) Changes of some enzyme activities of cucumber during ammonium toxicity. Plant Physiol 24: 248-258
  30. Wu JY, Wong K, Ho KP, Zhou LG (2005) Enhancement of saponin production in Panax ginseng cell culture by osmotic stress and nutrient feeding. Enzyme Microb Tech 36: 133-138 https://doi.org/10.1016/j.enzmictec.2004.07.010
  31. Xie JT, Wang A, Mehendale S, Wu J, Aung HH, Dey L, Qiu S, Yuan CS (2003) Antidiabetic effects of Gymnema yunnanese extract. Pharm Res 43: 323-329
  32. Zhang YH, Zhong JJ, Yu JT (1996) Enhancement of ginseng saponin production in suspension culture of Panax notoginseng. manipulation of medium sucrose. J Biotechnol 51: 49-56 https://doi.org/10.1016/0168-1656(96)01560-X
  33. Zhong JJ, Bai Y, Wang SJ (1996) Effects of plant growth regulatiors on cell growth and ginsenoside saponin production by suspension cultures of Panax quinquefolium. J Biotechnol 45: 227-234 https://doi.org/10.1016/0168-1656(95)00170-0
  34. Zhong JJ, Wang SJ (1998) Effects of nitrogen source on the production of ginseng saponin and polysaccharide by cell cultures of Panax quinquefolium. Proc Biochem 33: 671-675