Skip to main content

Advertisement

SpringerLink
Log in

Cell culture establishment and regulation of two phenylethanoid glycosides accumulation in cell suspension culture of desert plant Cistanche tubulosa

  • Original Article
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

Cistanche tubulosa is one of the most valuable desert medicinal plants, whose cell culture investigations have been rarely reported before. Phenylethanoid glycosides (PhGs) are its major components with a wide range of pharmacological activities. In this article, callus culture and cell suspension of C. tubulosa were established. Fleshy stems were found to be the most suitable explants for callus induction, and the optimal medium for induction was B5 solid medium supplemented with 0.8 g/L casein hydrolysate, 20 g/L sucrose, 2 mg/L naphthaleneacetic acid (NAA), and 1 mg/L 6-benzyladenine (6-BA). Based on qualitative and quantitative determination of two PhGs (echinacoside and acteoside) contents, the effects of carbon source concentration, precursor feeding, and elicitor treatments on cell growth and two PhGs accumulation in cell suspension cultures were investigated. Thirty g/L was the optimal initial sucrose concentration to obtain the high yield of biomass (9.29 g dry weight, DW) per liter cell suspension culture, echinacoside (12.14%, based on DW cells) and acteoside (2.17%). Precursor feeding also had a positive effect on PhGs accumulation. Feeding of precursor tyrosine (1 g/L) to the cell cultures increased the levels of echinacoside to 18.83% and acteoside to 2.92%, which were approximate 1.5 times of the corresponding levels in the control group. Methyl jasmonate (MJ) was the ideal elicitor for PhGs accumulations in C. tubulosa, particularly for eliciting acteoside production. The maximum echinacoside and acteoside contents reached 21.18 and 5.24% after 12 h of treatment with 200 µM MJ, respectively, which were approximate twofold higher than those in wild plant.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Ahmadi-Sakha S, Sharifi M, Niknam V (2016) Bioproduction of phenylethanoid glycosides by plant cell culture of Scrophularia striata Boiss.: from shake-flasks to bioreactor. Plant Cell Tissue Organ Cult 124:275–281

    Article  CAS  Google Scholar 

  • 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 25:613–620

    Article  PubMed  CAS  Google Scholar 

  • Bernard F, Moghbel N, Hassannejad S (2012) Treatment of licorice seeds with colchicine: changes in seedling DNA levels and anthocyanin and glycyrrhizic acid contents of derived callus cultures. Nat Prod Commun 7:1457–1460

    PubMed  CAS  Google Scholar 

  • Brooks CJW, Watson DG, Freer IM (1986) Elicitation of capsidiol accumulation in suspended callus-cultures of Capsicum annuum. Phytochemistry 25:1089–1092

    Article  CAS  Google Scholar 

  • Cheng XY, Guo B, Zhou HY, Ni W, Liu CZ (2005) Repeated elicitation enhances phenylethanoid glycosides accumulation in cell suspension cultures of Cistanche deserticola. Biochem Eng J 24:203–207

    Article  CAS  Google Scholar 

  • Creelman RA, Mullet JE (1997) Biosynthesis and action of jasmonates in plants. Annu Rev Plant Physiol Plant Mol Biol 48:355–381

    Article  PubMed  CAS  Google Scholar 

  • Draper J (1997) Salicylate, superoxide synthesis and cell suicide in plant defence. Trends Plant Sci 2:162–165

    Article  Google Scholar 

  • Ellis BE (1983) Production of hydroxyphenylethanol glycosides in suspension cultures of Syringa vulgaris. Phytochemistry 22:1941–1943

    Article  CAS  Google Scholar 

  • Ewelina P, Łukasz K, Ewa S, Marta Ż, Ewa B, Halina W (2015) Iridoid and phenylethanoid glycoside production and phenotypical changes in plants regenerated from hairy roots of rehmannia glutinosa, Libosch. Plant Cell Tissue Organ Cult 122:259–266

    Article  CAS  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  PubMed  CAS  Google Scholar 

  • Gu C, Yang X, Huang L (2016) Cistanches Herba: a neuropharmacology review. Front Pharmacol 7:289

    PubMed  PubMed Central  Google Scholar 

  • Hu GS, Jia JM, Kim DH (2014) Effects of feeding tyrosine and phenylalanine on the accumulation of phenylethanoid glycosides to Cistanche deserticola cell suspension culture. Chin J Nat Med 12:367–372

    PubMed  CAS  Google Scholar 

  • Jiang Y, Tu PF (2009) Analysis of chemical constituents in Cistanche species. J Chromatogr A 1216:1970–1979

    Article  PubMed  CAS  Google Scholar 

  • Kim SR, Kim Y, An G (1993) Identification of methyl jasmonate and salicylic acid response elements from the nopaline synthase (nos) promoter. Plant Physiol 103:97–103

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Largia MJV, Pothiraj G, Shilpha J, Ramesh M (2015) Methyl jasmonate and salicylic acid synergism enhances bacoside A content in shoot cultures of Bacopa monnieri (L.). Plant Cell Tissue Organ Cult 122:9–20

    Article  CAS  Google Scholar 

  • Li Z, Lin H, Gu L, Gao J, Tzeng CM (2016) Herba Cistanche (Rou Cong-Rong): one of the best pharmaceutical gifts of traditional Chinese medicine. Front Pharmacol 7:41

    PubMed  PubMed Central  Google Scholar 

  • Liu CZ, Cheng XY (2008) Enhancement of phenylethanoid glycosides biosynthesis in cell cultures of Cistanche deserticola by osmotic stress. Plant Cell Rep 27:357–362

    Article  PubMed  CAS  Google Scholar 

  • Liu JY, Guo ZG, Zeng ZL (2007) Improved accumulation of phenylethanoid glycosides by precursor feeding to suspension culture of Cistanche salsa. Biochem Eng J 33:88–93

    Article  CAS  Google Scholar 

  • Lu CT, Mei XG (2003) Improvement of phenylethanoid glycosides production by a fungal elicitor in cell suspension culture of Cistanche deserticola. Biotechnol Lett 25:1437–1439

    Article  PubMed  CAS  Google Scholar 

  • Lu DY, Zhang JY, Yang ZY, Liu HM, Li S, Wu BJ, Ma ZG (2013) Quantitative analysis of cistanches herba using high-performance liquid chromatography coupled with diode array detection and high-resolution mass spectrometry combined with chemometric methods. J Sep Sci 36:1945–1952

    Article  PubMed  CAS  Google Scholar 

  • Moriya A, Karasawa DE, Arima H, Deyama T, Ibe N, Kegasawa K, Usmanghani K (1995) Studies on the tissue culture of Cistanche plants: I. callus induction and shoot differentiation from the seed of Cistanche tubulosa wight. Plant Tissue Cult Lett 12:46–54

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Ouyang J, Wang XD, Zhao B, Wang YC (2003) Formation of phenylethanoid glycosides by Cistanche deserticola callus grown on solid media. Biotechnol Lett 25:223–225

    Article  PubMed  CAS  Google Scholar 

  • Ouyang J, Wang XD, Zhao B, Wang YC (2005) Enhanced production of phenylethanoid glycosides by precursor feeding to cell culture of Cistanche deserticola. Process Biochem 40:3480–3484

    Article  CAS  Google Scholar 

  • Ram M, Prasad KV, Singh SK, Hada BS, Kumar S (2013) Influence of salicylic acid and methyl jasmonate elicitation on anthocyanin production in callus cultures of Rosa hybrida, L. Plant Cell Tissue Organ Cult 113:459–467

    Article  CAS  Google Scholar 

  • Song YL, Song QQ, Li J, Zhang N, Zhao YF, Liu X, Jiang Y, Tu PF (2016) An integrated strategy to quantitatively differentiate chemome between Cistanche deserticola and C. tubulosa using high performance liquid chromatography-hybrid triple quadrupole-linear ion trap mass spectrometry. J Chromatogr A 1429:238–247

    Article  PubMed  CAS  Google Scholar 

  • Sudha G, Ravishankar GA (2002) Involvement and interaction of various signaling compounds on the plant metabolic events during defense response, resistance to stress factors, formation of secondary metabolites and their molecular aspects. Plant Cell Tissue Organ Cult 71:181–212

    Article  CAS  Google Scholar 

  • Wang T, Zhang X, Xie W Ma YC (2012) Cistanche deserticola. “Desert ginseng”: a review. Am J Chin Med 40:1123–1141

    Article  PubMed  CAS  Google Scholar 

  • Yan Y, Song QQ, Chen XJ, Li J, Li P, Wang YT, Liu TX, Song YL, Tu PF (2017) Simultaneous determination of components with wide polarity and content ranges in cistanche tubulosa using serially coupled reverse phase-hydrophilic interaction chromatography-tandem mass spectrometry. J Chromatogr A 1501:39–50

    Article  PubMed  CAS  Google Scholar 

  • Zhang JX, Ma LQ, Yu HS, Zhang H, Wang HT, Qin YF, Shi GL, Wang YN (2011) A tyrosine decarboxylase catalyzes the initial reaction of the salidroside biosynthesis pathway in Rhodiola sachalinensis. Plant Cell Rep 30:1443–1453

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

We are thankful to the financial supports by the National Natural Science Foundation of China (Grant No. 81402809) and independent project in Beijing University of Chinese Medicine (No. 2014-JYBZZ-JS-022).

Author information

Authors and Affiliations

  1. Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100029, People’s Republic of China

    Xiao Liu, Yaru Yan, Yuyu Liu, Ting Mo, Xiaohui Wang, Yuelin Song, Yunfang Zhao, Shepo Shi & Pengfei Tu

  2. School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing, 100191, People’s Republic of China

    Qingliang Chen & Pengfei Tu

  3. Ningxia Hui Medicine Research Institute, Yinchuan, 750021, People’s Republic of China

    Ting Mo

Authors
  1. Xiao Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yaru Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yuyu Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ting Mo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xiaohui Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yuelin Song
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Qingliang Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yunfang Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Shepo Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Pengfei Tu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

XL, YY, YL and TM carried out the experiments; XW and YZ performed statistical analysis of results; YS performed the HPLC and MS analyses work; QC developed seeds germination methods; XL drafted the manuscript; XL, SS and PT initiated the project and supervised the work throughout. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Xiao Liu, Shepo Shi or Pengfei Tu.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Communicated by Sergio J. Ochatt.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 4150 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, X., Yan, Y., Liu, Y. et al. Cell culture establishment and regulation of two phenylethanoid glycosides accumulation in cell suspension culture of desert plant Cistanche tubulosa. Plant Cell Tiss Organ Cult 134, 107–118 (2018). https://doi.org/10.1007/s11240-018-1404-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-018-1404-y

Keywords

Search

Navigation