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Plant growth regulator induced phytochemical and antioxidant variations in micropropagated and acclimatized Eucomis autumnalis subspecies autumnalis (Asparagaceae)

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Abstract

Eucomis species is a valuable plant with both medicinal and horticultural potential. The current study evaluated the role of plant growth regulator (PGR) on growth, phytochemicals, and antioxidant activity in Eucomis autumnalis subspecies autumnalis. Five cytokinins including topolins and benzyladenine (BA) at 2 µM in combination with varying (0–15 µM) concentrations of naphthalene acetic acid (NAA) were tested. In vitro regenerants were acclimatized in the greenhouse for 4 months. Highest number of shoots (9 shoots/explant) was observed with 15 µM NAA alone or when combined with BA. Acclimatized plants derived from the 15 µM NAA treatment had the highest number of roots, largest leaf area and biggest bulbs. While applied PGRs increased the iridoids and condensed tannins in the in vitro regenerants, total phenolics and flavonoids were higher in the PGR-free treatment. Among the in vitro regenerants, 5 µM NAA and 2 µM BA treatments produced the best antioxidant activity in the DPPH (55 %) and beta-carotene (88 %) test systems, respectively. A remarkable carry-over effect of the PGR was conspicuous in the phytochemical levels and antioxidant activity of the 4-month-old plants. In addition to the optimized micropropagation protocols, the current findings present a promising potential for manipulating the type and concentration of applied PGRs to improve phytochemical production and hence medicinal value in E. autumnalis subspecies autumnalis.

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References

  • Ahmad N, Khan MI, Ahmed S, Javed SB, Faisal M, Anis M, Rehman S, Umair SM (2013) Change in total phenolic content and antibacterial activity in regenerants of Vitex negundo L. Acta Physiol Plant 35:791–800

    CAS  Google Scholar 

  • Amâncio S, Rebordão J, Chaves M (1999) Improvement of acclimatization of micropropagated grapevine: photosynthetic competence and carbon allocation. Plant Cell Tissue Organ Cult 58:31–37

    Article  Google Scholar 

  • Amarowicz R, Pegg RB, Rahimi-Moghaddam P, Barl B, Weil JA (2004) Free-radical scavenging capacity and antioxidant activity of selected plant species from the Canadian prairies. Food Chem 84:551–562

    Article  CAS  Google Scholar 

  • Amoo SO, Van Staden J (2013) Influence of plant growth regulators on shoot proliferation and secondary metabolite production in micropropagated Huernia hystrix. Plant Cell Tissue Organ Cult 112:249–256

    Article  CAS  Google Scholar 

  • Amoo SO, Aremu AO, Van Staden J (2013) Shoot proliferation and rooting treatments influence secondary metabolite production and antioxidant activity in tissue culture-derived Aloe arborescens grown ex vitro. Plant Growth Regul 70:115–122

    Article  CAS  Google Scholar 

  • Amoo SO, Aremu AO, Moyo M, Szüčová L, Doležal K, Van Staden J (2014) Physiological effects of a novel aromatic cytokinin analogue in micropropagated Aloe arborescens and Harpagophytum procumbens. Plant Cell Tissue Organ Cult 116:17–26

    Article  CAS  Google Scholar 

  • Aremu AO, Bairu MW, Doležal K, Finnie JF, Van Staden J (2012a) Topolins: a panacea to plant tissue culture challenges? Plant Cell Tissue Organ Cult 108:1–16

    Article  CAS  Google Scholar 

  • Aremu AO, Bairu MW, Finnie JF, Van Staden J (2012b) Stimulatory role of smoke–water and karrikinolide on the photosynthetic pigment and phenolic contents of micropropagated ‘Williams’ bananas. Plant Growth Regul 67:271–279

    Article  CAS  Google Scholar 

  • Aremu AO, Bairu MW, Szüčová L, Doležal K, Finnie JF, Van Staden J (2012c) Assessment of the role of meta-topolins on in vitro produced phenolics and acclimatization competence of micropropagated ‘Williams’ banana. Acta Physiol Plant 34:2265–2273

    CAS  Google Scholar 

  • Aremu AO, Gruz J, Šubrtová M, Szüčová L, Doležal K, Bairu MW, Finnie JF, Van Staden J (2013) Antioxidant and phenolic acid profiles of tissue cultured and acclimatized Merwilla plumbea plantlets in relation to the applied cytokinins. J Plant Physiol 170:1303–1308

    Article  CAS  PubMed  Google Scholar 

  • Ault JR (1995) In vitro propagation of Eucomis autumnalis, E. comosa, and E. zambesiaca by twin-scaling. HortScience 30:1441–1442

    Google Scholar 

  • Bairu MW, Amoo SO, Van Staden J (2011) Comparative phytochemical analysis of wild and in vitro-derived greenhouse-grown tubers, in vitro shoots and callus-like basal tissues of Harpagophytum procumbens. S Afr J Bot 77:479–484

    Article  CAS  Google Scholar 

  • Cheesman L, Finnie JF, Van Staden J (2010) Eucomis zambesiaca Baker: factors affecting in vitro bulblet induction. S Afr J Bot 76:543–549

    Article  CAS  Google Scholar 

  • Coenen C, Lomax TL (1997) Auxin-cytokinin interactions in higher plants: old problems and new tools. Trends Plant Sci 2:351–356

    Article  CAS  PubMed  Google Scholar 

  • Coste A, Vlase L, Halmagyi A, Deliu C, Coldea G (2011) Effects of plant growth regulators and elicitors on production of secondary metabolites in shoot cultures of Hypericum hirsutum and Hypericum maculatum. Plant Cell Tissue Organ Cult 106:279–288

    Article  CAS  Google Scholar 

  • Doležal K, Popa I, Kryštof V, Spíchal L, Fojtíková M, Holub J, Lenobel R, Schmülling T, Strnad M (2006) Preparation and biological activity of 6-benzylaminopurine derivatives in plants and human cancer cells. Bioorg Med Chem 14:875–884

    Article  PubMed  Google Scholar 

  • Doležal K, Popa I, Hauserová E, Spíchal L, Chakrabarty K, Novák O, Kryštof V, Voller J, Holub J, Strnad M (2007) Preparation, biological activity and endogenous occurrence of N6-benzyladenosines. Bioorg Med Chem 15:3737–3747

    Article  PubMed  Google Scholar 

  • Dörnenburg H, Knorr D (1995) Strategies for the improvement of secondary metabolite production in plant cell cultures. Enzyme Microbial Technol 17:674–684

    Article  Google Scholar 

  • García-Pérez E, Gutiérrez-Uribe J, García-Lara S (2012) Luteolin content and antioxidant activity in micropropagated plants of Poliomintha glabrescens (Gray). Plant Cell Tissue Organ Cult 108:521–527

    Article  Google Scholar 

  • Gaspar T, Kevers C, Penel C, Greppin H, Reid D, Thorpe T (1996) Plant hormones and plant growth regulators in plant tissue culture. In Vitro Cell Dev Biol Plant 32:272–289

    Article  CAS  Google Scholar 

  • George EF, Hall MA, De Klerk G-J (2008) Plant growth regulators II: cytokinins, their analogues and antagonists. In: George EF, Hall MA, De Klerk G-J (eds) Plant propagation by tissue culture. Springer, Netherlands, pp 205–226

    Google Scholar 

  • Hazarika BN (2006) Morpho-physiological disorders in in vitro culture of plants. Sci Hortic 108:105–120

    Article  CAS  Google Scholar 

  • Hazarika RR, Chaturvedi R (2013) Establishment of dedifferentiated callus of haploid origin from unfertilized ovaries of tea (Camellia sinensis (L.) O. Kuntze) as a potential source of total phenolics and antioxidant activity. In Vitro Cell Dev Biol Plant 49:60–69

    Article  Google Scholar 

  • Huang D, Ou B, Prior RL (2005) The chemistry behind antioxidant capacity assays. J Agr Food Chem 53:1841–1856

    Article  CAS  Google Scholar 

  • Kozai T, Kubota C, Ryoung Jeong B (1997) Environmental control for the large-scale production of plants through in vitro techniques. Plant Cell Tissue Organ Cult 51:49-56

  • Liu CZ, Murch SJ, El-Demerdash M, Saxena PK (2004) Artemisia judaica L.: micropropagation and antioxidant activity. J Biotechnol 110:63–71

    Article  CAS  PubMed  Google Scholar 

  • Liu X-N, Zhang X-Q, Sun J-S (2007) Effects of cytokinins and elicitors on the production of hypericins and hyperforin metabolites in Hypericum sampsonii and Hypericum perforatum. Plant Growth Regul 53:207–214

    Article  CAS  Google Scholar 

  • Malá J, Máchová P, Cvrčková H, Karady M, Novák O, Mikulík J, Hauserová E, Greplová J, Strnad M, Doležal K (2009) Micropropagation of wild service tree (Sorbus torminalis [L.] Crantz): the regulative role of different aromatic cytokinins during organogenesis. J Plant Growth Regul 28:341–348

    Article  Google Scholar 

  • Masondo NA, Finnie JF, Van Staden J (2014) Pharmacological potential and conservation prospect of the genus Eucomis (Hyacinthaceae) endemic to southern Africa. J Ethnopharmacol 151:44–53

    Article  PubMed  Google Scholar 

  • Matkowski A (2008) Plant in vitro culture for the production of antioxidants—a review. Biotechnol Adv 26:548–560

    Article  CAS  PubMed  Google Scholar 

  • McCartan SA, Van Staden J (1995) In vitro propagation of the medicinal plant, Eucomis poleevansii N. E. Brown. J S Afr Soc Hortic Sci 5:73–75

    Google Scholar 

  • Moyo M, Ndhlala AR, Finnie JF, Van Staden J (2010) Phenolic composition, antioxidant, and acetylcholinesterase inhibitory activities of Sclerocarya birrea and Harpephyllum caffrum (Anacardiaceae) extracts. Food Chem 123:69–76

    Article  CAS  Google Scholar 

  • Moyo M, Aremu AO, Gruz J, Šubrtová M, Szüčová L, Doležal K, Van Staden J (2013) Conservation strategy for Pelargonium sidoides DC: phenolic profile and pharmacological activity of acclimatized plants derived from tissue culture. J Ethnopharmacol 149:557–561

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nordström A, Tarkowski P, Tarkowska D, Norbaek R, Ästot C, Dolezal K, Sandberd 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 21:8039–8044

    Article  Google Scholar 

  • Nunes JDM, Pinhatti AV, Rosa LMG, von Poser GL, Rech SB (2009) Roles of in vitro plantlet age and growing period in the phenolic constituent yields of acclimatized Hypericum polyanthemum. Env Exp Bot 67:204–208

    Article  CAS  Google Scholar 

  • Podlešáková K, Zalabák D, Čudejková M, Plíhal O, Szϋčová L, Doležal K, Spíchal L, Strnad M, Galuszka P (2012) Novel cytokinin derivatives do not show negative effects on root growth and proliferation in submicromolar range. PLoS ONE 7:e39293. doi:10.1371/journal.pone.0039293

    Article  PubMed Central  PubMed  Google Scholar 

  • Pospíšilová J, Synková H, Haisel D, Semorádová S (2007) Acclimation of plantlets to ex vitro conditions: effects of air humidity, irradiance, CO2 concentration and abscisic acid (a review). Acta Hortic 748:29–38

    Google Scholar 

  • Quiala E, Cañal M-J, Meijón M, Rodríguez R, Chávez M, Valledor L, de Feria M, Barbón R (2012) Morphological and physiological responses of proliferating shoots of teak to temporary immersion and BA treatments. Plant Cell Tissue Organ Cult 109:223–234

    Article  CAS  Google Scholar 

  • Ramachandra Rao S, Ravishankar GA (2002) Plant cell cultures: chemical factories of secondary metabolites. Biotechnol Adv 20:101–153

    Article  CAS  Google Scholar 

  • Strnad M, Hanus J, Vanek T, Kamínek M, Ballantine JA, Fussell B, Hanke DE (1997) Meta-topolin, a highly active aromatic cytokinin from poplar leaves (Populus × canadensis Moench., cv. Robusta). Phytochemistry 45:213–218

    Article  CAS  Google Scholar 

  • Szücová L, Spíchal L, Doležal K, Zatloukal M, Greplová J, Galuszka P, Krystof V, Voller J, Popa I, Massino FJ, Jørgensen J-E, Strnad M (2009) Synthesis, characterization and biological activity of ring-substituted 6-benzylamino-9-tetrahydropyran-2-yl and 9-tetrahydrofuran-2-ylpurine derivatives. Bioorg Med Chem 17:1938–1947

    Article  PubMed  Google Scholar 

  • Tarkowská D, Doležal K, Tarkowski P, Åstot C, Holub J, Fuksová K, Schmülling T, Sandberg G, Strnad M (2003) Identification of new aromatic cytokinins in Arabidopsis thaliana and Populus × canadensis leaves by LC-(+)ESI-MS and capillary liquid chromatography/frit-fast atom bombardment mass spectrometry. Physiol Plant 117:579–590

    Article  PubMed  Google Scholar 

  • Tarkowski P, Floková K, Václavíková K, Jaworek P, Raus M, Nordström A, Novák O, Doležal K, Šebela M, Frébortová J (2010) An improved in vivo deuterium labeling method for measuring the biosynthetic rate of cytokinins. Molecules 15:9214–9229

    Article  CAS  PubMed  Google Scholar 

  • Taylor JLS, Van Staden J (2001) In vitro propagation of Eucomis L’Herit species—plants with medicinal and horticultural potential. Plant Growth Regul 34:317–329

    Article  CAS  Google Scholar 

  • Valero-Aracama C, Kane M, Wilson S, Philman N (2010) Substitution of benzyladenine with meta-topolin during shoot multiplication increases acclimatization of difficult- and easy-to-acclimatize sea oats (Uniola paniculata L.) genotypes. Plant Growth Regul 60:43–49

    Article  CAS  Google Scholar 

  • Van Huylenbroeck JM, Piqueras A, Debergh PC (2000) The evolution of photosynthetic capacity and the antioxidant enzymatic system during acclimatization of micropropagated Calathea plants. Plant Sci 155:59–66

    Article  PubMed  Google Scholar 

  • Werbrouck SPO, Van der Jeugt B, Dewitte W, Prinsen E, Van Onckelen HA, Debergh PC (1995) The metabolism of benzyladenine in Spathiphyllum floribundum ‘Schott Petite’ in relation to acclimatisation problems. Plant Cell Rep 14:662–665

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

Financial support from the National Research Foundation (NAM), Claude Leon Foundation (AOA) and University of KwaZulu-Natal, South Africa is appreciated. Dr. K. Doležal and the team at Laboratory of Growth Regulators, Palacký University and Institute of Experimental Botany AS CR, Czech Republic for providing the topolins. Mrs. Alison Young (UKZN Botanical Garden) and Dr. C. Potgieter (Bews herbarium) assisted with the greenhouse experiments and plant identification, respectively. We are grateful to Drs. M. Moyo and S.O. Amoo for their valuable suggestions on the manuscript.

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  1. Research Centre for Plant Growth and Development, School of Life Sciences, University of KwaZulu-Natal Pietermaritzburg, Private Bag X01, Scottsville, 3209, South Africa

    Nqobile A. Masondo, Adeyemi O. Aremu, Jeffrey F. Finnie & Johannes Van Staden

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  1. Nqobile A. Masondo
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  2. Adeyemi O. Aremu
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  3. Jeffrey F. Finnie
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  4. Johannes Van Staden
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Correspondence to Johannes Van Staden.

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Communicated by M. J. Reigosa.

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Masondo, N.A., Aremu, A.O., Finnie, J.F. et al. Plant growth regulator induced phytochemical and antioxidant variations in micropropagated and acclimatized Eucomis autumnalis subspecies autumnalis (Asparagaceae). Acta Physiol Plant 36, 2467–2479 (2014). https://doi.org/10.1007/s11738-014-1619-4

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