Horticulture, Environment, and Biotechnology

, Volume 60, Issue 1, pp 125–134 | Cite as

Ginsenoside accumulation profiles in long- and short-term cell suspension and adventitious root cultures in Panax ginseng

  • Kim-Cuong Le
  • Cheol-Seung Jeong
  • Hyoshin Lee
  • Kee-Yoeup Paek
  • So-Young ParkEmail author
Research Report


In this study, we compared two in vitro culture systems, dedifferentiated cell suspension culture and differentiated adventitious root culture, using inoculum from long-term (20-year culture period) and short-term (1-year culture period) cultures, for the production of ginsenosides in wild ginseng (Panax ginseng). Increases in biomass and ginsenoside content were monitored in a 3-L bioreactor. The biomass in short-term cell suspension and adventitious root cultures increased at a faster rate than that in the long-term cultures, reaching a peak after 4 and 8 weeks of culture, respectively. In long term cultured materials, the biomass of cell suspension and adventitious root cultures decreased by 1.67- and 1.52-fold, respectively. Although the amount of ginsenosides in cell suspension and adventitious root cultures varied during the culture periods, the total ginsenoside content remained constant. These data suggest that both culture systems are advantageous for the production of different ginsenosides for pharmaceutical purposes.


Adventitious roots Biomass Bioreactor culture Cell suspension Ginsenoside Panax ginseng 



This study was funded by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through the Advanced Production Technology Development Program, funded by the Ministry of Agriculture, Food and Rural Affairs (Grant Number 315013-4), and Kim-Cuong Le was supported by the Brain Korea 21 (BK21) Plus program.

Author’s contribution

K-C L contributed to data acquisition and the writing of the manuscript. C-S J, H L, and K-Y P participated in data interpretation and revising of the manuscript to include important intellectual content. S-Y P made substantial contributions to data interpretation, revising of the manuscript, the conception, and design of this study.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.


  1. Ahmed EU, Hayashi T, Yazawa S (2004) Auxins increase the occurrence of leaf-colour variants in Caladium regenerated from leaf explants. Sci Hortic 100:153–159CrossRefGoogle Scholar
  2. Ahn SI, Kim SK, Yang BW, Lee ES, Kang CS, Hahm YT (2016) Analysis of ginsenosides and non-saponin components of red ginseng from landraces and new Varieties. Korean J Hortic Sci Technol 34:790–798Google Scholar
  3. Bairu MW, Aremu AO, Van Staden J (2011) Somaclonal variation in plants: causes and detection methods. J Plant Growth Regul 63:147–173CrossRefGoogle Scholar
  4. Bhatia S, Bera T (2015) Classical and nonclassical techniques for secondary metabolite production in plant cell culture. In: Bhatia S, Sharma K, Dahiya R, Tanmoy B (eds) Modern applications of plant biotechnology in pharmaceutical sciences. Elsevier, London, pp 231–291CrossRefGoogle Scholar
  5. Bhojwani SS, Razdan MK (1996) Plant tissue culture: theory and practice, a revised edition. Elsevier, AmsterdamGoogle Scholar
  6. Bolta Z, Baričevič D, Bohanec B, Andrenšek S (2000) A preliminary investigation of ursolic acid in cell suspension culture of Salvia officinalis. Plant Cell Tiss Org 62:57–63CrossRefGoogle Scholar
  7. Bourgaud F, Gravot A, Milesi S, Gontier E (2001) Production of plant secondary metabolites: a historical perspective. Plant Sci 161:839–851CrossRefGoogle Scholar
  8. Briskin DP (2007) Biotechnological methods for selection of high-yielding cell lines and production of secondary metabolites in medicinal plants. In: Kayser O, Quax WJ (eds) Medicinal plant biotechnology. From basic research to industrial applications. Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, pp 187–201Google Scholar
  9. Coppede JS, Pina ES, Paz TA, Fachin AL, Marins MA, Bertoni BW, França SC, Pereira AM (2014) Cell cultures of Maytenus ilicifolia Mart. are richer sources of quinone-methide triterpenoids than plant roots in natura. Plant Cell Tiss Organ Cult 118:33–43CrossRefGoogle Scholar
  10. Cui XH, Murthy HN, Paek KY (2014) Production of adventitious root biomass and bioactive compounds from Hypericum perforatum L. through large-scale bioreactor cultures. In: Paek KY, Murthy HN, Zhong JJ (eds) Production of biomass and bioactive compounds using bioreactor technology. Springer, Dordrecht, pp 251–284Google Scholar
  11. Dubrovina AS, Kiselev KV (2012) Effect of long-term cultivation on resveratrol accumulation in a high-producing cell culture of Vitis amurensis. Acta Physiol Plant 34:1101–1106CrossRefGoogle Scholar
  12. Fu C, Li L, Wu W, Li M, Yu X, Yu L (2012) Assessment of genetic and epigenetic variation during long-term Taxus cell culture. Plant Cell Rep 31:1321–1331CrossRefGoogle Scholar
  13. Garíca-Mateos MR, Gutiérrez RJM, Soto-Hernández RM, VillegasMonter A (2005) Alkaloids from several subcultures of Erythrina americana Miller calluses. Rev Chapingo Ser Hort 11:21–26CrossRefGoogle Scholar
  14. George EF, de Klerk GJ (2008) The components of plant tissue culture media I: macro- and micro-nutrients. In: George EF, Hall MA, de Klerk GJ (eds) Plant propagation by tissue culture, 3rd edn. Springer, Berlin, pp 65–114Google Scholar
  15. Hiai S, Oura H, Nakajima T (1976) Color reaction of some sapogenins and saponins with vanillin and sulfuric acid. Planta Med 29:116–122CrossRefGoogle Scholar
  16. Ho TT, Lee KJ, Lee JD, Bhushan S, Paek KY, Park SY (2017) Adventitious root culture of Polygonum multiflorum for phenolic compounds and its plot-scale production in 500 L-tank. Plant Cell Tiss Org 130:167–181CrossRefGoogle Scholar
  17. Kikowska M, Budzianowski J, Krawczyk A, Thiem M (2012) Accumulation of rosmarinic, chlorogenic and caffeic acids in in vitro cultures of Eryngium planum L. Acta Physiol Plant 34:2425–2433CrossRefGoogle Scholar
  18. Kim YJ, Zhang D, Yang DC (2015) Biosynthesis and biocoenological production of ginsenosides. Biotechnol Adv 33:717–735CrossRefGoogle Scholar
  19. Kim JK, Tabassum N, Uddin MR, Park SU (2016) Ginseng: a miracle sources of herbal and pharmacological uses. Orient Pharm Exp Med 16:243–250CrossRefGoogle Scholar
  20. Kiselev KV, Dubrovina AS, Shumakova OA (2013) DNA mutagenesis in 2-and 20-yr-old Panax ginseng cell cultures. In Vitro Cell Dev Biol-Plant 49:175–182CrossRefGoogle Scholar
  21. Langhansová L, Maršík P, Vaněk T (2005) Production of saponins from Panax ginseng suspension and adventitious root cultures. Biol Plant 49:463–465CrossRefGoogle Scholar
  22. Lee M, Phillips RL (1988) The chromosomal basis of somaclonal variation. Annu Rev Plant Physiol Plant Mol Biol 39:413–437CrossRefGoogle Scholar
  23. Lee JD, Le KC, Park YK, Murthy HN, Paek KY, Park SY (2018) Cell culture system versus adventitious root culture system in Asian and American ginseng: a collation. Plant Cell Tiss Org 132:295–302CrossRefGoogle Scholar
  24. Li L-Q, Fu C-H, Zhao C-F, Xia J, Wu W-J, Yu L-J (2009) Efficient extraction of RNA and analysis of gene expression in long-term Taxus cell culture using real-time RT-PCR. Z Naturforsch C 64:125–130CrossRefGoogle Scholar
  25. Loschiavo F, Pitto L, Giuliano G, Torti G, Nuti-Ronchi V, Marazziti D, Vergara R, Orselli S, Terzi M (1989) DNA methylation of embryogenic carrot cell cultures and its variations as caused by mutation, differentiation, hormones and hypomethylating drugs. Theor Appl Genet 77:325–331CrossRefGoogle Scholar
  26. Machczyńska J, Orłowska R, Mańkowski DR, Zimny J, Bednarek PT (2014) DNA methylation changes in triticale due to in vitro culture plant regeneration and consecutive reproduction. Plant Cell Tiss Org 119:289–299CrossRefGoogle Scholar
  27. Mancuso C, Santangelo R (2017) Panax ginseng and Panax quinquefolius: from pharmacology to toxicology. Food Chem Toxicol 107:362–372CrossRefGoogle Scholar
  28. Mather JP, Roberts PE (1998) Introduction to cell and tissue culture: theory and technique. Introductory cell and molecular biology techniques. Plenum Press, New YorkGoogle Scholar
  29. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Plant Physiol 15:473–497CrossRefGoogle Scholar
  30. Murthy HN, Georgiev MI, Kim YS, Jeong CS, Kim SJ, Park SY, Paek KY (2014a) Ginsenosides: prospective for sustainable biotechnological production. Appl Microbiol Biotechnol 98:6243–6254CrossRefGoogle Scholar
  31. Murthy HN, Kim YS, Jeong CS, Kim SJ, Zhong JJ, Paek KY (2014b) Production of ginsenosides from adventitious root cultures of Panax ginseng. In: Paek KY, Murthy HN, Zhong JJ (eds) Production of biomass and bioactive compounds using bioreactor technology. Springer, Dordrecht, pp 625–651Google Scholar
  32. Murthy HN, Kim YS, Park SY, Paek KY (2014c) Hypericins: biotechnological production from cell and organ cultures. Appl Microbiol Biotechnol 98:9187–9198CrossRefGoogle Scholar
  33. Nagella P, Murthy HN (2014) Production of withanolides from cell and organ cultures of Withania somnifera (L.) Dunal. In: Paek KY, Murthy HN, Zhong JJ (eds) Production of biomass and bioactive compounds using bioreactor technology. Springer, Dordrecht, pp 285–316Google Scholar
  34. Paek KY, Murthy HN, Hahn EJ, Zhong JJ (2009) Large scale culture of ginseng adventitious roots for production of ginsenosides. In: Zhong JJ, Bai FW, Zhang W (eds) Biotechnology in China I. Advances in Biochemical Engineering/Biotechnology, vol 113. pp 151–176Google Scholar
  35. Park SY, Paek KY (2014) Bioreactor culture of shoots and somatic embryos of medicinal plants for production of bioactive compounds. In: Paek KY, Murthy HN, Zhong JJ (eds) Production of biomass and bioactive compounds using bioreactor technology. Springer, Dordrecht, pp 337–368Google Scholar
  36. Park JD, Rhee DK, Lee YH (2005) Biological activities and chemistry of saponins from Panax ginseng CA Meyer. Phytochem Rev 4:159–175CrossRefGoogle Scholar
  37. Park SY, Park JH, Kim HS, Lee CY, Lee HJ, Kang KS, Kim CE (2017) Systems-level mechanisms of action of Panax ginseng: a network pharmacological approach. J Ginseng Res 42:98–106CrossRefGoogle Scholar
  38. Patil RA, Kolewe ME, Roberts SC (2013) Cellular aggregation is a key parameter associated with long term variability in paclitaxel accumulation in Taxus suspension cultures. Plant Cell Tissue Organ Cult 112:303–310CrossRefGoogle Scholar
  39. Qu JG, Zhang W, Yu XJ, Jin MF (2005) Instability of anthocyanin accumulation in Vitis vinifera L. var. Gamay Freaux suspension cultures. Biotechnol Bioprocess Eng 10:155–161CrossRefGoogle Scholar
  40. Schumann A, Claus D, Gerth A (2014) In vitro production of Digitalis purpurea biomass using temporary immersion cultures. In: Paek KY, Murthy HN, Zhong JJ (eds) Production of biomass and bioactive compounds using bioreactor technology. Springer, Dordrecht, pp 417–428Google Scholar
  41. Shi W, Wang Y, Li J, Zhang H, Ding L (2007) Investigation of ginsenosides in different parts and ages of Panax ginseng. Food Chem 102:664–668CrossRefGoogle Scholar
  42. Sierra MI, van der Heijden R, van der Leer T, Verpoorte R (1992) Stability of alkaloid production in cell suspension cultures of Tabernaemontana divaricata during long-term subculture. Plant Cell Tissue Organ Cult 28:59–68CrossRefGoogle Scholar
  43. Skirvin RM, Chu MC, Mann ML, Young H, Sullivan J, Fermanian T (1986) Stability of tissue culture medium pH as a function of autoclaving, time, and cultured plant material. Plant Cell Rep 5:292–294CrossRefGoogle Scholar
  44. Thanh NT, Murthy HN, Paek KY (2014) Ginseng cell culture for production of ginsenosides. In: Paek KY, Murthy HN, Zhong JJ (eds) Production of biomass and bioactive compounds using bioreactor technology. Springer, Dordrecht, pp 121–142Google Scholar
  45. Thorpe T, Stasolla C, Yeung EC, de Klerk GJ, Roberts A, George EF (2008) The components of plant tissue culture media II: organic additions, osmotic and pH effects, and support systems. In: George EF, Hall MA, de Klerk GJ (eds) Plant propagation by tissue culture, 3rd edn. Springer, Berlin, pp 115–174Google Scholar
  46. Trejo-Tapia G, Balcazar-Aguilar JB, Martínez-Bonfil B, Salcedo-Morales G, Jaramillo-Flores M, Arenas-Ocampo ML, JiménezAparicio A (2008) Effect of screening and subculture on the production of betaxanthins in Beta vulgaris L. var. ‘Dark Detroit’ callus culture. Innov Food Sci Emerg Technol 9:32–36CrossRefGoogle Scholar
  47. Wang JW, Wu JY (2010) Tanshinone biosynthesis in Salvia miltiorrhiza and production in plant tissue cultures. Appl Microbiol Biotechnol 88:437–449CrossRefGoogle Scholar
  48. Whitmer S, Canel C, van der Heijden R, Verpoorte R (2003) Long-term instability of alkaloid production by stably transformed cell lines of Catharanthus roseus. Plant Cell Tiss Org 74:73–80CrossRefGoogle Scholar
  49. Yeoman MM, Yeoman CL (1996) Manipulating secondary metabolism in cultured plant cells. New Phytol 134:553–569CrossRefGoogle Scholar
  50. Zobayed SMA (2006) Aeration in plant tissue culture-engineering aspects of vessel design. In: Gupta SD, Ibaraki Y (eds) Plant tissue culture engineering. Springer, Dordrecht, pp 313–327Google Scholar

Copyright information

© Korean Society for Horticultural Science 2018

Authors and Affiliations

  • Kim-Cuong Le
    • 1
  • Cheol-Seung Jeong
    • 1
    • 2
  • Hyoshin Lee
    • 3
  • Kee-Yoeup Paek
    • 1
    • 2
  • So-Young Park
    • 1
    Email author
  1. 1.Division of Animal, Horticulture and Food SciencesChungbuk National UniversityCheongjuRepublic of Korea
  2. 2.WellGreen Co.CheongjuRepublic of Korea
  3. 3.Department of Forest Genetic ResourcesNational Institute of Forest ScienceSuwonRepublic of Korea

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