Skip to main content
SpringerLink
Log in

Improvement of shoot proliferation and comparison of secondary metabolites in shoot and callus cultures of Phlomis armeniaca by LC-ESI-MS/MS analysis

  • Plant Tissue Culture
  • Published:
In Vitro Cellular & Developmental Biology - Plant Aims and scope Submit manuscript

Abstract

Phlomis armeniaca Willd. is a medicinal plant in the Lamiaceae family endemic to Turkey. The present study describes efficient plant regeneration and callus induction protocols for P. armeniaca and compares phenolic profiles, total phenol and flavonoid contents, and free radical scavenging activity of in vitro-derived tissues. Stem node explants from germinated seedlings were cultured on Murashige and Skoog medium (MS) supplemented with 75 plant growth regulator (PGR) combinations. The highest shoot number per explant, frequency of shoot proliferation, and frequency of highly proliferated, green, compact callus were obtained on MS medium containing 0.25 mg L−1 thidiazuron (TDZ) and 0.25 mg L−1 indole-3-acetic acid (IAA). The best root formation was on MS basal medium (control). Methanol extract of leaves obtained from regenerants contained higher total phenol and flavonoid contents than the callus extract. The callus extract showed stronger free radical scavenging activity than leaves with IC50 [concentration inhibiting 50% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical] values of 4.30 ± 0.08 and 2.21 ± 0.04 mg g−1 dry weight in leaves and callus, respectively. Apigenin, caffeic acid, p-coumaric acid, luteolin, rutin hydrate, vanillic acid, ferulic acid, salicylic acid, sinapic acid, and chlorogenic acid were detected by liquid chromatography–electrospray ionization multistage tandem mass spectrometry (LC-ESI-MS/MS) analysis in in vitro-grown leaves and callus tissue. Rutin hydrate, p-coumaric acid, and vanillic acid were found at approximately tenfold higher levels in callus than in leaves. This new micropropagation protocol, the first for P. armeniaca, could be used in industrial production for new herbal tea and germplasm conservation.

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

Figure 1.
Figure 2.
Figure 3.

Similar content being viewed by others

References

  • Babbar N, Oberoi HS, Uppal DS, Patil RT (2011) Total phenolic content and antioxidant capacity of extracts obtained from six important fruit residues. Food Res Intern 44:391–396

    Article  CAS  Google Scholar 

  • Baytop T (1999) Therapy with medicinal plants in Turkey; today and in future (in Turkish). Istanbul University Press, Istanbul

    Google Scholar 

  • Brand-Williams W, Cuvelier ME, Berset C (1995) Use of a free radical method to evaluate antioxidant activity. LWT––Food Sci Technol 28:25–30

    CAS  Google Scholar 

  • Calis I, Kirmizibekmez H, Ersoz T, Saracoglu I, Donmez Ali A, Mitova M, Handjieva N, Popov S (2002) Iridoid, phenylethanoid flavonoid glycosides from Phlomis sintenisii. Turk Acta Pharm Turc 44:195–203

    CAS  Google Scholar 

  • Chang C, Yang M, Wen H, Chern J (2002) Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Anal 10:178–182

    CAS  Google Scholar 

  • Cook NC, Samman S (1996) Flavonoids—chemistry, metabolism, cardioprotective effects, and dietary sources. Nutr Biochem 7:66–76

    Article  CAS  Google Scholar 

  • Dalar A, Konczak I (2014) Cichorium intybus from eastern Anatolia: phenolic composition, antioxidant and enzyme inhibitory activities. Ind Crop Prod 60:79–85

    Article  CAS  Google Scholar 

  • De Gyves EM, Sparks CA, Fieldsend AF, Lazzeri PA, Jones HD (2001) High frequency of adventitious shoot regeneration from commercial cultivars of evening primrose (Oenothera spp.) using thidiazuron. Ann Appl Biol 138:329–332

    Article  Google Scholar 

  • Demirci F, Guven K, Demirci B, Dadandi MY, Baser KHC (2008) Antibacterial activity of two Phlomis essential oils against food pathogens. Food Control 19:1159–1164

    Article  CAS  Google Scholar 

  • Dewanto V, Wu X, Adom KK, Liu RH (2002) Thermal processing enhances the nutritional value of tomatoes by increasing total antioxidant activity. J Agr Food Chem 50:3010–3014

    Article  CAS  Google Scholar 

  • Ersoz T, Ivancheva S, Akbay P, Sticher O, Calis I (2001) Iridoid and phenylethanoid glycosides from Phlomis tuberosa L. Z Naturforsch C 56:695–698

    Article  CAS  PubMed  Google Scholar 

  • Fernandes de Oliveira AM, Sousa Pinheiro L, Souto Pereira CK, Neves Matias W, Albuquerque Gomes R, Souza Chaves O, Vanderlei de Souza MF, Nóbrega de Almeida R, Simões de Assis T (2012) Total phenolic content and antioxidant activity of some Malvaceae family species. Antioxidants 1:33–43

    Article  CAS  PubMed  Google Scholar 

  • Gençay A (2007) Cizre (Şırnak)' nin etnobotanik özellikleri. Yüzüncü Yıl Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, Van, Turkey

  • Gonçalves S, Romano A (2013) In vitro culture of lavenders (Lavandula spp.) and the production of secondary metabolites. Biotechnol Adv 31:166–174

    Article  PubMed  Google Scholar 

  • Grzegorczyk-Karolak I, Kuzma L, Wysokinska H (2015) The effect of cytokinins on shoot proliferation, secondary metabolite production and antioxidant potential in shoot cultures of Scutellaria alpina. Plant Cell Tissue Organ Cult 122:699–708

    Article  CAS  Google Scholar 

  • Gülçin I, Oktay M, Kireçci E, Küfrevioğlu İÖ (2003) Screening of antioxidant and antimicrobial activities of anise (Pimpinella anisum L.) seed extracts. Food Chem 83:371–382

    Article  Google Scholar 

  • Huetteman CA, Preece JE (1993) Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tissue Organ Cult 33:105–119

    Article  CAS  Google Scholar 

  • Kabouche A, Kabouche Z, Seguin E, Tillequin F, Bruneau C (2005) A phenylethanoid glycoside and flavonoids from Phlomis crinita (Cav.) Lamiaceae. Biochem Syst Ecol 33:813–816

    Article  CAS  Google Scholar 

  • Karakas FP, Turker AU (2013) An efficient in vitro regeneration system for Bellis perennis L. and comparison of phenolic contents of field-grown and in vitro-grown leaves by LC-MS/MS. Ind Crop Prod 48:162–170

    Article  CAS  Google Scholar 

  • Karakas FP, Yildirim AB, Bayram R, Yavuz ZP, Gepdiremen A, Turker AU (2015) Antiproliferative activity of some medicinal plants on human breast and hepatocellular carcinoma cell lines and their phenolic contents. Trop J Pharm Res 14:1787–1795

    Article  CAS  Google Scholar 

  • Limem-Ben Amor I, Boubaker J, Ben Sgaier M, Skandrani I, Bhouri W, Neffati A, Kilani S, Bouhlel I, Ghedira K, Chekir-Ghedira L (2009) Phytochemistry and biological activities of Phlomis species. J Ethnopharmacol 125:183–202

    Article  CAS  PubMed  Google Scholar 

  • Mahendran G, Narmatha Bai V (2014) Micropropagation, antioxidant properties and phytochemical assessment of Swertia corymbosa (Griseb.) Wight ex C. B. Clarke: a medicinal plant. Acta Physiol Plant 36:589–603

    Article  CAS  Google Scholar 

  • Mallon R, Rodriguez-Oubina J, Gonzalez ML (2011) Shoot regeneration from in vitro-derived leaf and root explants of Centaurea ultreiae. Plant Cell Tissue Organ Cult 106:523–530

    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 

  • Nordström A, Tarkowski P, Tarkowska D, Norbaek R, Åstot C, Dolezal K, Sandberg G (2004) Auxin regulation of cytokinin biosynthesis in Arabidopsis thaliana: a factor of potential importance for auxin–cytokinin-regulated development. Proc Natl Acad Sci USA 101:8039–8044

    Article  PubMed  PubMed Central  Google Scholar 

  • Pehlivan Karakas F, Sahin G, Turker A (2016) Enhancement of direct shoot regeneration and determination of bioactive secondary metabolites in leaves of Galega officinalis L. Turk J Biol (in press). doi:10.3906/biy-1603-70

    Google Scholar 

  • Pokorny J (1991) Natural antioxidants for food use. Trends Food Sci Technol 2:223–227

    Article  CAS  Google Scholar 

  • Saracoglu I, Inome M, Calis I, Ogihara Y (1995) Studies on constituents with cytotoxic and cytostatic activity of 2 Turkish medicinal-plants Phlomis armeniaca and Scutellaria salviifolia. Biol Pharm Bull 18:1396–1400

    Article  CAS  PubMed  Google Scholar 

  • Sarikurkcu C, Uren MC, Tepe B, Cengiz M, Kocak MS (2015) Phlomis armeniaca: phenolic compounds, enzyme inhibitory and antioxidant activities. Ind Crop Prod 78:95–101

    Article  CAS  Google Scholar 

  • Shahidi F (2000) Antioxidants in food and food antioxidants. Nahrung 44:158–163

    Article  CAS  PubMed  Google Scholar 

  • Turker AU, Yıldırım AB (2013) Evaluation of antibacterial and antitumor activities of some Turkish endemic plants. Trop J Pharm Res 12:1003–1010

    Article  Google Scholar 

  • Turker A, Yucesan B, Gurel E (2009) In vitro regeneration of Achillea millefolium L. from shoot-tips and root segments of seedlings. J Plant Biochem Biotechnol 18:61–69

    Article  Google Scholar 

  • Uysal A, Gunes E, Sarikurkcu C, Celik H, Durak Y, Uren MC (2016) New prospective materials for chemoprevention: three Phlomis. Br J Pharm Res 10(3):1–13

    Google Scholar 

  • Verpoorte R, Memelink J (2002) Engineering secondary metabolite production in plants. Curr Opin Biotechnol 13:181–187

    Article  CAS  PubMed  Google Scholar 

  • Yalcin FN, Ersoz T, Akbay P, Calis I, Donmez AA, Sticher O (2003) Iridoid and phenylpropanoid glycosides from Phlomis samia, P. monocephala and P. carica. Turk J Chem 27:295–305

    CAS  Google Scholar 

  • Yasar S, Fakir H, Erbas S (2010) Gas chromatographic (GC-GC/MS) analysis of essential oil of Phlomis armeniaca Willd. from mediterranean region in Turkey. Asian J Chem 22:2887–2890

    CAS  Google Scholar 

  • Yucesan B, Turker AU, Gurel E (2007) TDZ-induced high frequency plant regeneration through multiple shoot formation in witloof chicory (Cichorium intybus L.). Plant Cell Tissue Organ Cult 91:243–250

    Article  CAS  Google Scholar 

  • Yumrutas O, Saygıdeger SD (2012) Determination of antioxidant and antimutagenic activities of Phlomis armeniaca and Mentha pulegium. J App Pharm Sci 2:36–40

    Google Scholar 

Download references

Acknowledgement

This study was supported by grants from the Abant Izzet Baysal University Research Foundation (project no. 2013.03.01.625).

Author information

Authors and Affiliations

  1. Department of Field Crops, Faculty of Agriculture and Natural Sciences, Abant Izzet Baysal University, Bolu, Turkey

    Fatma Pehlivan Karakas

  2. Department of Biology, Faculty of Science and Art, Abant Izzet Baysal University, Bolu, Turkey

    Fatma Pehlivan Karakas & Arzu Ucar Turker

Authors
  1. Fatma Pehlivan Karakas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Arzu Ucar Turker
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Fatma Pehlivan Karakas.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interest.

Additional information

Editor: Rakhi Chaturvedi

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Karakas, F.P., Turker, A.U. Improvement of shoot proliferation and comparison of secondary metabolites in shoot and callus cultures of Phlomis armeniaca by LC-ESI-MS/MS analysis. In Vitro Cell.Dev.Biol.-Plant 52, 608–618 (2016). https://doi.org/10.1007/s11627-016-9792-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11627-016-9792-3

Keywords

Search

Navigation