Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 137, Issue 1, pp 101–113 | Cite as

Improvement of bioactive saponin accumulation in adventitious root cultures of Panax vietnamensis via culture periods and elicitation

  • Nguyen Thi Nhat Linh
  • Le Kim Cuong
  • Ho Thanh Tam
  • Hoang Thanh Tung
  • Vu Quoc Luan
  • Vu Thi Hien
  • Nguyen Hoang Loc
  • Duong Tan NhutEmail author
Original Article


Saponin compounds released by Panax vietnamensis into the rhizosphere play critical roles in drug discovery. This study investigated the growth and saponin accumulation of P. vietnamensis adventitious root. Results showed the root growth and productivity of three-saponins (MR2, Rb1, and Rg1) were increased highest on the modified MS medium containing 7 mg/L IBA and 0.5 mg/L BA at 56 days. The root culture divided into three phases: (1) Growth phase 14–56 culture-days; (2) Stationary phase 56–70 days; and (3) Decline phase after 70 days. Saponin accumulation was significantly increased in the treatments of three plant-hormone-elicitors (JA, ABA, and SA), and two cell-wall elicitors (YE and CHN), but five elicitors inhibited formation lateral roots. Although contents of Rg1 (0.32%), Rb1 (0.85%), and MR2 (2.83%) were highest in JA treatment at 0.48 mM, 0.24 mM, and 0.96 mM JA respectively, maximum biomass as well as saponins productivity was observed in 150 mg/L YE. In addition, combination of YE and SA showed effective lower than the solitary YE treatment. The results suggest that adventitious root biomass of P. vietnamensis developed effectively in the modified MS medium after 56 days, and the culture could be scale-up in bioreactor 20 L.


Saponin Adventitious root culture Elicitors Plant growth regulators P. vietnamensis 



This study was supported by Tay Nguyen Institute for Scientific Research (VAST) and The Research Centre of Ginseng and Medicinal Materials in Vietnam.

Author contributions

NTNL and DTN conceived of the presented idea. NTNL, LKC, HTT, HTT, VQL, and VTH developed the theory and performed the computations. NTNL verified the analytical methods. DTN and NHL encouraged NTNL, LKC, HTT, HTT, VQL, and VTH to investigate the development of adventitious root and supervised the findings of this work. All authors discussed the results and contributed to the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. Ali MB, Yu KW, Hahn EJ, Paek KY (2006) 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 25(6):613–620. Google Scholar
  2. Chang K, Chen M, Zeng L, Lan X, Wang Q, Liao Z (2014) Abscisic acid enhanced ajmalicine biosynthesis in hairy roots of Rauvolfia verticillata by upregulating expression of the MEP pathway genes. Russ J Plant Physiol 61(1):136–140Google Scholar
  3. Cho HY, Son SY, Rhee HS, Yoon SY, Lee-Parsons CW, Park JM (2008) Synergistic effects of sequential treatment with methyl jasmonate, salicylic acid and yeast extract on benzophenanthridine alkaloid accumulation and protein expression in Eschscholtzia californica suspension cultures. J Biotechnol 135(1):117–122. Google Scholar
  4. Duc NM, Kasai R, Ohtani K, Ito A, Nguyen TN, Yamasaki K, Tanaka O (1994) Saponins from Vietnamese Ginseng, Panax vietnamensis Ha et Grushv. Collected in Central Vietnam. III. Chem Pharm Bull 42(3):634–640. Google Scholar
  5. Duncan DB (1955) Multiple range and multiple F tests. Biometrics 11(1):1–42. Google Scholar
  6. Gregory LR (2009) Mechanical induction of lateral root initiation. The Pennsylvania State University, 108Google Scholar
  7. Guo D, Liang J, Li L (2009) Abscisic acid (ABA) inhibition of lateral root formation involves endogenous ABA biosynthesis in Arachis hypogaea L. Plant Growth Regul 58(2):173–179. Google Scholar
  8. Han JY, In JG, Kwon YS, Choi YE (2010) Regulation of ginsenoside and phytosterol biosynthesis by RNA interferences of squalene epoxidase gene in Panax ginseng. Phytochemistry 71(1):36–46. Google Scholar
  9. Hasanloo T, Sepehrifar R, Rahnama H, Shams MR (2009) Evaluation of the yeast-extract signaling pathway leading to silymarin biosynthesis in milk thistle hairy root culture. World J Microbiol Biotechnol 25(11):1901–1909. Google Scholar
  10. Ho TT, Lee KJ, Lee JD, Bhushan S, Paek KY, Park SY (2017) Adventitious root culture of Polygonum multiflorum for phenolic compounds and its pilot-scale production in 500 L-tank. Plant Cell Tissue Organ Cult 130(1):167–181. Google Scholar
  11. Ho TT, Lee JD, Jeong CS, Paek KY, Park SY (2018) Improvement of biosynthesis and accumulation of bioactive compounds by elicitation in adventitious root cultures of P. multiflorum. Appl Microbiol Biotechnol 102(1):199–209. Google Scholar
  12. Hu FX, Zhong JJ (2007) Role of jasmonic acid in alteration of ginsenoside heterogeneity in elicited cell cultures of Panax notoginseng. J Biosci Bioeng 104(6):513–516. Google Scholar
  13. Hu X, Zhang W, Fang J, Cai W, Tang Z (2002) Chitosan treatment raises the accumulation of saponin and the transcriptional Level of genes encoding the key enzymes of saponin synthesis in cultured Panax ginseng cells. Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao 28(6):485–490Google Scholar
  14. Hu X, Neill SJ, Fang J, Cai W, Tang Z (2004) Mitogen-activated protein kinases mediate the oxidative burst and saponin synthesis induced by chitosan in cell cultures of Panax ginseng. Sci China C Life Sci 47(4):303–312Google Scholar
  15. Jeong JJ, Van Le TH, Lee SY, Eun SH, Nguyen MD, Park JH, Kim DH (2015) Anti-inflammatory effects of vina-ginsenoside R2 and majonoside R2 isolated from Panax vietnamensis and their metabolites in lipopolysaccharide-stimulated macrophages. Int Immunopharmacol 28(1):700–706. Google Scholar
  16. Kang SM, Min JY, Kim YD, Kang YM, Park DJ, Jung HN, Kim SW, Choi MS (2006) Effects of methyl jasmonate and salicylic acid on the production of bilobalide and ginkgolides in cell cultures of Ginkgo biloba. In Vitro Cell Dev Biol Plant 42(1):44–49. Google Scholar
  17. Kim YS, Hahn EJ, Murthy HN, Paek KY (2004) Adventitious root growth and ginsenoside accumulation in Panax ginseng cultures as affected by methyl jasmonate. Biotechnol Lett 26(21):1619–1622. Google Scholar
  18. Marsik P, Langhansova L, Dvorakova M, Cigler P, Hruby M, Vanek T (2014) Increased ginsenosides production by elicitation of in vitro cultivated Panax ginseng adventitious roots. Med Aromat Plants 3:1–5. Google Scholar
  19. Mejia-Teniente L, de Dalia Duran-Flores F, Chapa-Oliver AM, Torres-Pacheco I, Cruz-Hernandez A, Gonzalez-Chavira MM, Ocampo-Velazquez RV, Guevara-Gonzalez RG (2013) Oxidative and molecular responses in Capsicum annuum L. after hydrogen peroxide, salicylic acid and chitosan foliar applications. Int J Mol Sci 14(5):10178–10196. Google Scholar
  20. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497. Google Scholar
  21. Nhut DT, Huy NP, Chien HX, Luan TC, Thao LB (2012) In vitro culture of petiole longitudinal thin cell layer explants of vietnamese ginseng (Panax vietnamensis Ha et Grushv.) and preliminary analysis of saponin content. Int J Appl Biol Pharm 3(3):178–190Google Scholar
  22. Pirian K, Piri K (2013) Influence of yeast extract as a biotic elicitor on noradrenaline production in hairy root culture of Portulaca oleracea L. Int J Plant Prod 4(11):2960–2964Google Scholar
  23. Rahimi S, Devi BSR, Khorolragchaa A, Kim YJ, Kim JH, Jung SK, Yang DC (2014) Effect of salicylic acid and yeast extract on the accumulation of jasmonic acid and sesquiterpenoids in Panax ginseng adventitious roots. Russ J Plant Physiol 61(6):811–817. Google Scholar
  24. Schenk RU, Hildebrandt A (1972) Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Can J Res 50(1):199–204Google Scholar
  25. Sheng DF, Zhang YL (2013) Effects of ABA on tanshinones accumulation of Salvia miltiorrhiza hairy root. Zhong Yao Cai 36(3):354–358Google Scholar
  26. Wang W, Zhao ZJ, Xu Y, Qian X, Zhong JJ (2006) Efficient induction of ginsenoside biosynthesis and alteration of ginsenoside heterogeneity in cell cultures of Panax notoginseng by using chemically synthesized 2-hydroxyethyl jasmonate. Appl Microbiol Biotechnol 70(3):298–307. Google Scholar
  27. Yu KW, Gao WY, Hahn EJ, Paek KY (2001) Effects of macro elements and nitrogen source on adventitious root growth and ginsenoside production in ginseng (Panax ginseng C. A. Meyer). J Plant Biol 44(4):179–184. Google Scholar
  28. Yu KW, Gao W, Hahn EJ, Paek KY (2002) Jasmonic acid improves ginsenoside accumulation in adventitious root culture of Panax ginseng C.A. Meyer. Biochem Eng J 11(2):211–215. Google Scholar
  29. Zhao SJ, Li CY, Qian YC, Luo XP, Zhang X, Wang XS, Kang BY (2004) Induction of hairy roots of Panax ginseng and studies on suitable culture condition of ginseng hairy roots. Sheng Wu Gong Cheng Xue Bao 20(2):215–220Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Nguyen Thi Nhat Linh
    • 1
    • 2
  • Le Kim Cuong
    • 1
  • Ho Thanh Tam
    • 1
  • Hoang Thanh Tung
    • 1
  • Vu Quoc Luan
    • 1
  • Vu Thi Hien
    • 1
  • Nguyen Hoang Loc
    • 2
  • Duong Tan Nhut
    • 1
    Email author
  1. 1.Tay Nguyen Institute for Scientific ResearchVietnam Academy of Science and TechnologyHanoiVietnam
  2. 2.Hue University of SciencesHueVietnam

Personalised recommendations