Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
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The improvement of ginsenoside accumulation in Panax ginseng as a result of γ-irradiation

  • Kim, Dong Sub (Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute) ;
  • Song, Mira (Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute) ;
  • Kim, Sun-Hee (Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute) ;
  • Jang, Duk-Soo (Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute) ;
  • Kim, Jin-Baek (Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute) ;
  • Ha, Bo-Keun (Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute) ;
  • Kim, Sang Hoon (Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute) ;
  • Lee, Kyung Jun (Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute) ;
  • Kang, Si-Yong (Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute) ;
  • Jeong, Il Yun (Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute)
  • Received : 2012.05.16
  • Accepted : 2013.05.06
  • Published : 2013.07.15
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Abstract

In this study, gamma rays were used to irradiate embryogenic calli induced from cotyledon explants of Panax ginseng Meyer. After the embryogenic calli were irradiated, they were transferred to adventitious roots using an induction medium; next, mutated adventitious root (MAR) lines with a high frequency of adventitious root formations were selected. Two MAR lines (MAR 5-2 and MAR 5-9) from the calli treated with 50 Gy of gamma rays were cultured on an $NH_4NO_3$-free Murashige and Skoog medium with indole-3-butyric acid 3 mg/L. The expression of genes related to ginsenoside biosynthesis was analyzed using reverse transcription polymerase chain reaction with RNA prepared from native ginseng (NG), non-irradiated adventitious root (NAR) and 2 MAR lines. The expression of the squalene epoxidase and dammarenediol synthase genes was increased in the MAR 5-2 line, whereas the phytosterol synthase was increased in the MAR 5-9 line. The content and pattern of major ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1) were analyzed in the NG, NAR, and 2 MAR lines (MAR 5-2 and MAR 5-9) using TLC and HPLC. In the TLC analysis, the ginsenoside patterns in the NG, NAR, and 2 MAR lines were similar; in contrast, the MAR 5-9 line showed strong bands of primary ginsenosides. In the HPLC analysis, compared with the NG, one new type of ginsenoside was observed in the NAR and 2 MAR lines, and another new type of ginsenoside was observed in the 2 MAR lines irradiated with gamma rays. The ginsenoside content of the MAR 5-9 line was significantly greater in comparison to the NG.

Keywords

References

  1. Gillis CN. Panax ginseng pharmacology: a nitric oxide link? Biochem Pharmacol 1997;54:1-8. https://doi.org/10.1016/S0006-2952(97)00193-7
  2. Keum YS, Park KK, Lee JM, Chun KS, Park JH, Lee SK, Kwon H, Surh YJ. Antioxidant and anti-tumor promoting activities of the methanol extract of heat-processed ginseng. Cancer Lett 2000;150:41-48. https://doi.org/10.1016/S0304-3835(99)00369-9
  3. Sticher O. Getting to the root of ginseng. Chemtech 1998;28:26-32.
  4. Kim YK, Guo Q, Packer L. Free radical scavenging activity of red ginseng aqueous extracts. Toxicology2002;172:149-156. https://doi.org/10.1016/S0300-483X(01)00585-6
  5. Vogler BK, Pittler MH, Ernst E. The efficacy of ginseng: a systematic review of randomised clinical trials. Eur J Clin Pharmacol 1999;55:567-575. https://doi.org/10.1007/s002280050674
  6. Shibata S. Chemistry and cancer preventing activities of ginseng saponins and some related triterpenoid compounds. J Korean Med Sci 2001;16 Suppl:S28-S37.
  7. Kiefer D, Pantuso T. Panax ginseng. Am Fam Physician 2003;68:1539-1542.
  8. Kalinowska M, Zimowski J, Paczkowski C, Wojciechowski ZA. The formation of sugar chains triterpenoid saponins and glycoalkaloids. Phytochem Rev 2005;4:237-257. https://doi.org/10.1007/s11101-005-1422-3
  9. Shibata S, Tanaka O, Soma K, Ando T, Iida Y, Nakamura H. Studies on saponins and sapogenins of ginseng. The structure of panaxatriol. Tetrahedron Lett 1965;42:207-213.
  10. Palazon J, Cusido RM, Bonfill M, Mallol A, Moyano E, Morales C, Pinol MT. Elicitation of different Panax ginseng transformed root phenotypes for an improved ginsenoside production. Plant Physiol Biochem 2003;41: 1019-1025. https://doi.org/10.1016/j.plaphy.2003.09.002
  11. Kim MK, Lee BS, In JG, Sun H, Yoon JH, Yang DC. Comparative analysis of expressed sequence tags (ESTs) of ginseng leaf. Plant Cell Rep 2006;25:599-606. https://doi.org/10.1007/s00299-005-0095-0
  12. Shi W, Wang Y, Li J, Zhang H, Ding L. Investigation of ginsenosides in different parts and ages of Panx ginseng. Food Chem 2007;102:664-668. https://doi.org/10.1016/j.foodchem.2006.05.053
  13. Furuya Y, Yoshikawa T, Orihar Y, Oda H. Studies of the culture conditions for Panax ginseng cells in jar fermentors. J Nat Prod 1984;47:70-75. https://doi.org/10.1021/np50031a008
  14. Yu KW, Gao W, Hahn EJ, Paek KY. Jasmonic acid improves ginsenoside accumulation in adventitious root culture of Panax ginseng C.A. Meyer. Biochem Eng J 2002;11:211-215. https://doi.org/10.1016/S1369-703X(02)00029-3
  15. Woo SS, Song JS, Lee JY, In DS, Chung HJ, Liu JR, Choi DW. Selection of high ginsenoside producing ginseng hairy root lines using targeted metabolic analysis. Phytochemistry 2004;65:2751-2761. https://doi.org/10.1016/j.phytochem.2004.08.039
  16. Ali MB, Yu KW, Hahn EJ, Paek KY. Methyl jasmonate and salicylic acid elicitation induces ginsenosides accumulation, enzymatic and non-enzymatic antioxidant in suspension culture Panax ginseng roots in bioreactors. Plant Cell Rep 2006;25:613-620. https://doi.org/10.1007/s00299-005-0065-6
  17. Asaka I, Li I, Hirotani M, Asada Y, Furuya T. Production of ginsenoside saponins by culturing ginseng (Panax ginseng) embryogenic tissues in bioreactors. Biotechnol Lett 1993;15:1259-1264. https://doi.org/10.1007/BF00130308
  18. Choi SM, Son SH, Yun SR, Kwon OW, Seon JH, Paek KY. Pilot-scale culture of adventitious roots of ginseng in a bioreactor system. Plant Cell Tissue Organ Cult 2000; 62:187-193. https://doi.org/10.1023/A:1006412203197
  19. Sivakumar G, Yu KW, Hahn EJ, Paek KY. Optimization of organic nutrients for ginseng hairy roots production in large-scale bioreactors. Curr Sci 2005;89:641-649.
  20. Thanh NT, Murthy HN, Yu KW, Hahn EJ, Paek KY. Methyl jasmonate elicitation enhanced synthesis of ginsenoside by cell suspension cultures of Panax ginseng in 5-l balloon type bubble bioreactors. Appl Microbiol Biotechnol 2005;67:197-201. https://doi.org/10.1007/s00253-004-1759-3
  21. Larkin PJ, Scowcroft WR. Somaclonal variation: a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 1981;60:197-214. https://doi.org/10.1007/BF02342540
  22. Kaeppler SM, Kaeppler HF, Rhee Y. Epigenetic aspects of somaclonal variation in plants. Plant Mol Biol 2000;43:179-188.
  23. Nakano M, Amano J, Watanabe Y, Nomizu T, Suzuki M, Mizunashi K, Mori S, Kuwayama S, Han DS, Saito H et al. Morphological variation in Tricyrtis hirta plants regenerated from heavy ion beam-irradiated embryogenic calluses. Plant Biotechnol 2010;27:155-160. https://doi.org/10.5511/plantbiotechnology.27.155
  24. Kovalchuk I, Molinier J, Yao Y, Arkhipov A, Kovalchuk O. Transcriptome analysis reveals fundamental differences in plant response to acute and chronic exposure to ionizing radiation. Mutat Res 2007;624:101-113. https://doi.org/10.1016/j.mrfmmm.2007.04.009
  25. Rakwal R, Agrawal GK, Shibato J, Imanaka T, Fukutani S, Tamogami S, Endo S, Sahoo SK, Masuo Y, Kimura S. Ultra low-dose radiation: stress responses and impacts using rice as a grass model. Int J Mol Sci 2009;10:1215-1225. https://doi.org/10.3390/ijms10031215
  26. Kim DS, Lee IS, Jang CS, Kang SY, Seo YW. Characterization of the altered anthranilate synthase in 5-methyltryptophan-resistant rice mutants. Plant Cell Rep 2005;24:357-365. https://doi.org/10.1007/s00299-005-0943-y
  27. Lee GJ, Chung SJ, Park IS, Lee JS, Kim JB, Kim DS, Kang SY. Variation in the phenotypic features and transcripts of color mutants of chrysanthemum (Dendranthema grandiflorum) derived from gamma ray mutagenesis. J Plant Biol 2008;51:418-423. https://doi.org/10.1007/BF03036063
  28. Arnold NP, Barthakur NN, Tanguay M. Mutagenic effects of acute gamma irradiation on miniature roses: target theory approach. HortScience 1998;33:127-129.
  29. Das A, Gosal SS, Sidhu JS, Dhaliwal HS. Induction of mutations for heat tolerance in potato by using in vitro culture and radiation. Euphytica 2000;114:205-209. https://doi.org/10.1023/A:1003965724880
  30. Patade VY, Suprasanna P. Radiation induced in vitro mutagenesis for sugarcane improvement. Sugar Tech 2008;10:14-19. https://doi.org/10.1007/s12355-008-0002-4
  31. Bermudez-Caraballoso I, Garcia LR, Veitia N, Torres D, Pardon Y, Romero C, Orellana P. Mutant plantains (Musa spp.) with height reduction obtained by in vitro mutagenesis. Euphytica 2010;176:105-112. https://doi.org/10.1007/s10681-010-0233-9
  32. Kim DS, Kim SY, Jeong IY, Kim JB, Lee GJ, Kang SY, Kim W. Improvement of ginsenoside production by Panax ginseng adventitious roots induced by $\gamma$-irradiation. Biol Plant 2009;53:408-414. https://doi.org/10.1007/s10535-009-0079-y
  33. Court WA, Hendel GJ, Elmi J. Reversed-phase high-performance liquid chromatography determination of ginsenosides of Panax quinquefolium. J Chromatogr A 1996;775:11-17.
  34. Bonfill M, Cusido RM, Palazon J, Teresa Pinol M, Morales C. Influence of auxins on organogenesis and ginsenoside production in Panax ginseng calluses. Plant Cell Tissue Organ Cult 2002;68:73-78. https://doi.org/10.1023/A:1012996116836
  35. Han JY, Kwon YS, Yang DC, Jung YR, Choi YE. Expression and RNA interference-induced silencing of the dammarenediol synthase gene in Panax ginseng. Plant Cell Physiol 2006;47:1653-1662. https://doi.org/10.1093/pcp/pcl032
  36. Abe I, Rohmer M, Prestwich GD. Enzymatic cyclization of squalene and oxidosqualene to sterols and triterpenes. Chem Rev 1993;93:2189-2206. https://doi.org/10.1021/cr00022a009
  37. Han JY, In JG, Kwon YS, Choi YE. Regulation of ginsenoside and phytosterol biosynthesis by RNA interferences of squalene epoxidase gene in Panax ginseng. Phytochemistry 2010;71:36-46. https://doi.org/10.1016/j.phytochem.2009.09.031
  38. Han JY, Jung SJ, Kim SW, Kwon KS, Yi MJ, Yi JS, Cho YE. Induction of adventitious roots and analysis of ginsenoside content and the genes involved in triterpene biosynthesis in Panax ginseng. J Plant Biol 2006;49:26-33. https://doi.org/10.1007/BF03030785
  39. Han JY, Choi YE. Rapid induction of Agrobacterium tumefaciens-mediated transgenic roots directly from adventitious roots in Panax ginseng. Plant Cell Tissue Organ Cult 2009;96:143-149. https://doi.org/10.1007/s11240-008-9470-1
  40. Wu J, Zhong JJ. Production of ginseng and its bioactive components in plant cell culture: current technological and applied aspects. J Biotechnol 1999;68:89-99. https://doi.org/10.1016/S0168-1656(98)00195-3
  41. Langhansova L, Marsik P, Vanek T. Production of saponins from Panax ginseng suspension and adventitious root cultures. Biol Plant 2005;49:463-465. https://doi.org/10.1007/s10535-005-0030-9
  42. Kim DS, Lee KJ, Yim WC, Kim JB, Ha BK, Kim SH, Kang SY. Transcriptional network analysis of the tryptophan-accumulating rice mutant during grain filling. Mol Genet Genomics 2012;287:699-709. https://doi.org/10.1007/s00438-012-0712-x

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