Enhanced expression of ginsenoside biosynthetic genes and in vitro ginsenoside production in elicited Panax sikkimensis (Ban) cell suspensions
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Dual metabolite, i.e., ginsenoside and anthocyanin, co-accumulating cell suspensions of Panax sikkimensis were subjected to elicitation with culture filtrates of Serratia marcescens (SD 21), Bacillus subtilis (FL11), Trichoderma atroviridae (TA), and T. harzianum (TH) at 1.25% and 2.5% v/v for 1- and 3-week duration. The fungal-derived elicitors (TA and TH) did not significantly affect biomass accumulation; however, bacterial elicitors (SD 21 and FL11), especially SD 21, led to comparable loss in biomass growth. In terms of ginsenoside content, differential responses were observed. A maximum of 3.2-fold increase (222.2 mg/L) in total ginsenoside content was observed with the use of 2.5% v/v TH culture filtrate for 1 week. Similar ginsenoside accumulation was observed with the use of 1-week treatment with 2.5% v/v SD 21 culture filtrate (189.3 mg/L) with a 10-fold increase in intracellular Rg2 biosynthesis (31 mg/L). Real-time PCR analysis of key ginsenoside biosynthesis genes, i.e., FPS, SQS, DDS, PPDS, and PPTS, revealed prominent upregulation of particularly PPTS expression (20–23-fold), accounting for the observed enhancement in protopanaxatriol ginsenosides. However, none of the elicitors led to successful enhancement in in vitro anthocyanin accumulation as compared to control values.
KeywordsPanax sikkimensis Cell suspension Ginsenoside Trichoderma Elicitation
The authors are grateful to the Director, CSIR-CIMAP, for the infrastructure and the lab facilities provided for the studies. TB also acknowledges the award of a Senior Research Fellowship (SRF) granted by the University Grants Commission, India.
Compliance with ethical standards
This article does not contain any studies with human participants or animals performed by any of the authors.
Conflict of interest
The authors declare that they have no conflict of interest.
- Biswas T (2016) Elicitation of in vitro secondary metabolite production and its transcript expression profiling in Panax species. PhD Thesis, Jawarharlal Nehru University, Delhi, IndiaGoogle Scholar
- Biswas T, Ajayakumar PV, Mathur AK, Mathur A (2015a) Solvent-based extraction optimization for efficient ultrasonication-assisted ginsenoside recovery from Panax quinquefolius and P. sikkimensis cell suspension lines. Natural Product Res 29(13):1256–1263. https://doi.org/10.1080/14786419.2015.1024119 CrossRefGoogle Scholar
- Biswas T, Kalra A, Mathur AK, Lal RK, Singh M, Mathur A (2016) Elicitors’ influenced differential ginsenoside production and exudation into medium with concurrent Rg3/Rh2 panaxadiol induction in Panax quinquefolius cell suspensions. Appl Microbiol Biotechnol 100(11):4909–4922. https://doi.org/10.1007/s00253-015-7264-z CrossRefPubMedGoogle Scholar
- Condori J, Sivakumar G, Hubstenberger J, Dolan MC, Sobolev VS, Medina-Bolivar F (2010) Induced biosynthesis of resveratrol and the prenylated stilbenoids arachidin-1 and arachidin-3 in hairy root cultures of peanut: effects of culture medium and growth stage. Plant Physiol Biochem 48(5):310–318. https://doi.org/10.1016/j.plaphy.2010.01.008 CrossRefPubMedGoogle Scholar
- Court WE (2000) Introduction to ginseng. In: Hardman R (ed) Ginseng—the genus Panax Hardwood. Academic Publishers, Netherlands, pp 1–11Google Scholar
- Li J, Liu S, Wang J, Li J, Liu D, Li J, Gao W (2016) Fungal elicitors enhance ginsenosides biosynthesis, expression of functional genes as well as signal molecules accumulation in adventitious roots of Panax ginseng CA Mey. J Biotechnol 239:106–114. https://doi.org/10.1016/j.jbiotec.2016.10.011 CrossRefPubMedGoogle Scholar
- Mañero F, Gutiérrezv J, Algar E, Martín Gómez MS, Saco Sierra MD, Solano BR (2012) Elicitation of secondary metabolism in Hypericum perforatum by rhizosphere bacteria and derived elicitors in seedlings and shoot cultures. Pharm Biol 50(10):1201–1209. https://doi.org/10.3109/13880209.2012.664150 CrossRefPubMedGoogle Scholar
- Mathur A, Gangwar A, Mathur AK, Sangwan RS, Jain DC (2002) A procedure for the development of an anthocyanin producing callus line of Panax sikkimensis (Indian species of ginseng) US PATENT 6368860.Google Scholar
- Mathur A, Mathur AK, Gangwar A, Verma P, Sangwan RS (2010) Anthocyanin production in a callus line of Panax sikkimensis Ban. In Vitro Cell Develop Biol-Plant 46(1):13–21. https://doi.org/10.1007/s11627-009-9253-3
- Mehta JK, Haridasan K (1992) The ginsengs in Arunachal Pradesh. Arunachal Forest news 10:56–58Google Scholar
- Paek KY, Murthy HN, Hahn EJ, Zhong JJ (2009) Large scale culture of ginseng adventitious roots for production of ginsenosides. Adv Biochem Engg Biotechnol 113:151–176Google Scholar
- Ryu CM, Choi HK, Lee CH, Murphy JF, Lee JK, Kloepper JW (2013) Modulation of quorum sensing in acyl-homoserine lactone-producing or -degrading tobacco plants leads to alteration of induced systemic resistance elicited by the rhizobacterium Serratia marcescens 90-166. Plant Pathol J 29(2):182–192. https://doi.org/10.5423/PPJ.SI.11.2012.0173 CrossRefPubMedPubMedCentralGoogle Scholar
- Singh RK, Chaudhary BD (1979) Biometrical methods in quantitative genetic analysis. Kalyani Publishers, New DelhiGoogle Scholar
- Smolenskaya IN, Reshetnyak OV, Nosov AV, Zoriniants SE, Chaiko AL, Smirnova YN, Nosov AM (2007) Ginsenoside production, growth and cytogenetic characteristics of sustained Panax japonicus var. repens cell suspension culture. Biol Plant 51(2):235–241. https://doi.org/10.1007/s10535-007-0047-3 CrossRefGoogle Scholar
- Thanh NT, Murthy HN, Yu KW, Hahn EJ, Paek KY (2005) Methyl jasmonate elicitation enhanced synthesis of ginsenoside by cell suspension cultures of Panax ginseng in 5-l balloon type bubble bioreactors. Appl Microbiol Biotechnol 67(2):197–201. https://doi.org/10.1007/s00253-004-1759-3 CrossRefPubMedGoogle Scholar
- Thanh NT, Ket NV, Yoeup PK (2007) Effect of medium composition on biomass and ginsenoside production in cell suspension culture of Panax vietnamensis Ha et Grushv. VNU J Sci Nat Sci Technol 23:269–274Google Scholar
- Xu MJ, Dong JF, Zhu MY (2005) Nitric oxide mediates the fungal elicitor-induced hypericin production of Hypericum perforatum cell suspension cultures through a jasmonic-acid-dependent signal pathway. Plant Physiol 139(2):991–998. https://doi.org/10.1104/pp.105.066407 CrossRefPubMedPubMedCentralGoogle Scholar