Advertisement

Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 131, Issue 2, pp 347–357 | Cite as

The effect of cytokinins on growth, phenolics, antioxidant and antimicrobial potential in liquid agitated shoot cultures of Knautia sarajevensis

  • Erna KaralijaEmail author
  • Sanja Ćavar Zeljković
  • Petr Tarkowski
  • Edina Muratović
  • Adisa Parić
Original Article

Abstract

Endemic plants are under constant threat of extinction and micropropagation protocols that can provide not only mechanisms for their revitalisation, but also a possibility for potential sources of new biologically active substances. This study describes the very first use of the liquid culture system for in vitro Knautia sarajevensis shoot multiplication, production of biomass, and an increase in biological activities of the extracts. Murashige and Skoog media containing various concentrations of cytokinins (6-benzyladenine, zeatin, and kinetin) were used to establish agitated shoot cultures of this endemic plant species and to evaluate their effect on shoot morphology. HPLC analysis of phenolic compounds show that the main metabolite in all extracts was salicylic acid with accumulation of rosmarinic and 4-hydroxybenzoic acid for some treatments. The richest sources of phenolics were shoots cultivated in media containing zeatin, which also had high influence on biomass production. Analysis of antioxidant and antimicrobial potential suggests that this plant could have beneficial biological activities. Methanol extracts of shoots cultivated in media containing 2.0 mg/L 6-benzyladenine were moderately active against Staphylococcus aureus and Bacillus spizizeni. DPPH assay showed radical scavenging activity of shoot cultures with IC50 10–90 µg/mL.

Keywords

Antibacterial activity Antioxidant activity In vitro cultures Phenolics HPLC 

Abbreviations

4HBA

4-Hydroxybenzoic acid

AP

Antioxidant potential

BA

6-Benzyladenine

CA

Caffeic acid

CGA

Chlorogenic acid

CHR

Chrysin

DPPH

2,2-Diphenyl-1-picrylhydrazyl radical

SD

Standard deviation

DW

Dry weight

FA

Ferulic acid

GA

Gallic acid

GAL

Galangin

HPLC

High-performance liquid chromatography

KIN

Kinetin

MS

Murashige and Skoog

MYR

Myricetin

PGR

Plant growth regulator

PIN

Pinocembrin

QUE

Quercetin

RA

Rosmarinic acid

SaA

Salicylic acid

SiA

Sinapic acid

SPA

Specific peroxidase activity

SyA

Syringic acid

TPC

Total protein content

VA

Vanillic acid

ZEA

Zeatin

Notes

Acknowledgements

This research was supported by Grant No. LO1204 (Sustainable development of research in the Centre of the Region Haná) from the National Program of Sustainability I, MEYS.

Author contributions

EK performed all the in vitro culture and analysis of growth, total phenolics, peroxidase activity and DPPH, antimicrobial potential, statistical analysis as well as the writing of the manuscript. SCZ and PT developed HPLC method for separation and quantification of phenolic compounds. They performed construction of calibration curves, analysis of plant extracts, and calculation of the content of each particular phenolic compound in extracts. EM participated in field work by collecting the seeds and identification of plant species used for this research and in statistical analysis of the data. AP supervised the in vitro culture work.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Akram M, Aftab F (2015) Effect of cytokinins on in vitro seed germination and changes in chlorophyll and soluble protein contents of teak (Tectona grandis L.). Biochem Physiol 4:166. doi: 10.4172/2168-9652.1000166 CrossRefGoogle Scholar
  2. Alali FQ, Amrine CSM, El-Elimat T, Alkofahi A, Tawaha K, Gharaibah M, Swanson SM, Falkinham JO, Cabeza M, Sánchez A, Figueroa M (2014) Bioactive withanolides from Withania obtusifolia. Phytochem Lett 9:96–101CrossRefGoogle Scholar
  3. Albayrak S, Aksoy A, Sagdic O, Hamzaoglu E (2010) Compositions, antioxidant and antimicrobial activities of Helichrysum (Asteraceae) species collected from Turkey. Food Chem 119(1):114–122CrossRefGoogle Scholar
  4. Allen DE, Hatfield G (2004) Medicinal plants in folk tradition. Timber Press, PortlandGoogle Scholar
  5. Barba-Espin G, Diaz-Vivancos P, Clemente-Moreno MJ, Albacete A, Faize L, Faize M, Pérez-Alfocea F, Hernández JA (2010) Interaction between hydrogen peroxide and plant hormones during germination and the early growth of pea seedlings. Plant Cell Environ 33(6):981–994CrossRefPubMedGoogle Scholar
  6. Beck GM, Maly K, Bijelćić Ž (1974) Knautia. In: Čović B (ed) Flora Bodnae et Herzegovinae, IV Sympetalae, vol 3. Zemaljski muzej Bosne i Hercegovine, Sarajevo, p 75Google Scholar
  7. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72(1–2):248–254CrossRefPubMedGoogle Scholar
  8. Gadzovska S, Maury S, Delaunay A, Spasenoski M, Joseph C, Hagege D (2007) Jasmonic acid elicitation of Hypericum perforatum L. cell suspensions and effects on the production of phenylpropanoids and naphtodianthrones. Plant Cell Tissue Organ Cult 89(1):1–13CrossRefGoogle Scholar
  9. González LF, Rojas MC, Perez FJ (1999) Diferulate and lignin formation is related tobiochemical differences of wall-bound peroxidases. Phytochemistry 50(5):711–717CrossRefGoogle Scholar
  10. Grzegorczyk-Karolak I, Rytczak P, Bielecki S, Wysokińska H (2015) The influence of liquid systems for shoot multiplication, secondary metabolite production and plant regeneration of Scutellaria alpina. Plant Cell Tissue Organ Cult 122(3):699–708CrossRefGoogle Scholar
  11. Grzegorczyk-Karolak I, Rytczak P, Bielecki S, Wysokińska H (2017) The influence of liquid systems for shoot multiplication, secondary metabolite production and plant regeneration of Scutellaria alpina. Plant Cell Tissue Organ Cult 128(2):479–486CrossRefGoogle Scholar
  12. Hendrawati O, Hille J, Woerdenbag HJ, Quax WJ, Kayser O (2012) In vitro regeneration of wild chervil (Anthriscus sylvestris L.). In Vitro Cell Dev Biol Plant 48(3):355–361CrossRefPubMedGoogle Scholar
  13. Karalija E, Ćavar Zeljković S, Tarkowski P, Muratović E, Parić A (2017) Media composition effects seed dormancy, apical dominance and phenolic profile of Knautia sarajevensis (Dipsacaceae), Bosnian endemic. Acta Bot Croat. doi: 10.1515/botcro-2017-0011
  14. Kasparavičienė G, Ramanauskienė K, Savickas A, Velžienė S, Kalvėnienė Z, Kazlauskienė D, Ragažinskienė O Ivanauskas K (2013) Evaluation of total phenolic content and antioxidant activity of different Rosmarinus officinalis L. ethanolic extracts. Biologija 59(1):39–44Google Scholar
  15. Kikowska M, Kędziora I, Krawczyk A, Thiem B (2015) Methyl jasmonate, yeast extract and sucrose stimulate phenolic acids accumulation in Eryngium planum L. shoot cultures. Acta Biochim Pol 62(2):197–2000CrossRefPubMedGoogle Scholar
  16. Kuhlmann A, Röhl C (2006) Phenolic antioxidant compounds produced by in vitro. Cultures of Rosemary (Rosmarinus officinalis.) and their anti-inflammatory effect on lipopolysaccharide-activated microglia. Pharm Biol 44(6):401–410CrossRefGoogle Scholar
  17. Lou Z, Wang H, Li J, Zhu S, Lu W, Ma C (2011) Effect of simultaneous ultrasonic/microwave assisted extraction on the antioxidant and antibacterial activities of burdock leaves. J Med Plant Res 5(22):5370–5377Google Scholar
  18. Matkowski A (2008) Plant in vitro culture for the production of antioxidants: a review. Biotechnol Adv 26(6):548–560CrossRefPubMedGoogle Scholar
  19. Mattalia G, Quave CL, Pieroni A (2013) Traditional uses of wild food and medicinal plants among Brigasc, Kyé, and Provençal communities on the Western Italian Alps. Genet Resour Crop Evol 60(2):587–603CrossRefGoogle Scholar
  20. McFarland J (1907) The nephelometer: an instrument for estimating the number of bacteria in suspensions used for calculating the opsonic index and for vaccines. JAMA 49(14):1176–1178CrossRefGoogle Scholar
  21. Meda A, Lamien CE, Romito M, Millogo J, Nacoulma OG (2005) Determination of the total phenolic, flavonoid and proline contents in Burkina Fasan honey, as well as their radical scavenging activity. Food Chem 91(3):571–577CrossRefGoogle Scholar
  22. Moore DM (1980) Dipsacaceae. In: Tutin TG, Heywood VH, Burges NA, Moore DM, Valentine DH, Walters SM, Webb DA (eds) Flora Europea, volume 4: plantaginaceae to compositae. University Press, Cambridge, p 56.Google Scholar
  23. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497CrossRefGoogle Scholar
  24. Murthy HN, Lee EJ, Paek KY (2014) Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell Tissue Organ Cult 118(1):1–16CrossRefGoogle Scholar
  25. Ordonez AAL, Gomez JD, Vattuone MA (2006) Antioxidant activities of Sechium edule (Jacq.) Swartz extracts. Food Chem 97(3):452–458CrossRefGoogle Scholar
  26. Ramirez-Estrada K, Vidal-Limon H, Hidalgo D, Moyano E, Golenioswki M, Cusidó RM, Palazon J (2016) Elicitation, an effective strategy for the biotechnological production of bioactive high-added value compounds in plant cell factories. Molecules 21(2):182CrossRefPubMedGoogle Scholar
  27. Raomai S, Kumaria S, Kehie M, Tandon P (2015) Plantlet regeneration of Paris polyphylla Sm. via thin cell layer culture and enhancement of steroidal saponins in mini-rhizome cultures using elicitors. Plant Growth Regul 75(1):341–353CrossRefGoogle Scholar
  28. Saher S, Piqueras A, Hellin E, Olmos E (2004) Hyperhydricity in micropropagated carnation shoots: the role of oxidative stress. Physiol Plant 120(1):152–161CrossRefPubMedGoogle Scholar
  29. Sakamoto K, Iida K, Sawamura K, Hajiro K, Asada Y, Yoshikawa T, Furuya T (1994) Anthocyanin production in cultured cells of Aralia cordata Thunb. Plant Cell Tissue Organ Cult 36(1):21–26CrossRefGoogle Scholar
  30. Savio, L. E. B., Astarita LV, Santarém ER (2012) Secondary metabolism in micropropagated Hypericum perforatum L. grown in non-aerated liquid medium. Plant Cell Tissue Organ Cult 108(3):465–472CrossRefGoogle Scholar
  31. Shani E, Ben-Gera H, Shleizer-Burko S, Burko Y, Weiss D, Ori N (2010) Cytokinin regulates compound leaf development in tomato. Plant Cell 22(10):3206–3217CrossRefPubMedPubMedCentralGoogle Scholar
  32. Takayama S, Akita M (2005) Practical aspects of bioreactor application in mass propagation of plants. In: Liquid culture systems for in vitro plant propagation. Springer, Dordrecht, pp 61–78CrossRefGoogle Scholar
  33. Tawaha K, Alali FQ, Gharaibeh M, Mohammad M, El-Elimat T (2007) Antioxidant activity and total phenolic content of selected Jordanian plant species. Food Chem 104(4):1372–1378CrossRefGoogle Scholar
  34. Tosun M, Ercisli S, Sengul M, Ozer H, Polat T, Ozturk E (2009) Antioxidant properties and total phenolic content of eight Salvia species from Turkey. Biol Res 42(2):175–181CrossRefPubMedGoogle Scholar
  35. Von Wettstein D, Jende-Strid B, Ahrenst-Larsen B, Sørensen JA (1977) Biochemical mutant in barley renders chemical stabilization of beer superfluous. Carlsberg Res Commun 42(5):341–351CrossRefGoogle Scholar
  36. Wawrosch C, Benda E, Kopp B (2009) An improved 2-step liquid culture system for efficient in vitro shoot proliferation of Sundew (Drosera rotundifolia L.). Sci Pharm 77(4):827–836Google Scholar
  37. Wolfe K, Wu X, Liu RH (2003) Antioxidant activity of apple peels. J Agric Food Chem 51(3):609–614CrossRefPubMedGoogle Scholar
  38. Xia D, Wu X, Shi J, Yang Q, Zhang Y (2011) Phenolic compounds from the edible seeds extract of Chinese Mei (Prunus mume Sieb. et Zucc) and their antimicrobial activity. LWT Food Sci Technol 44(1):347–349CrossRefGoogle Scholar
  39. Zaheer M, Giri CC (2015) Multiple shoot induction and jasmonic versus salicylic acid driven elicitation for enhanced andrographolide production in Andrographis paniculata. Plant Cell Tissue Organ Cult 122(3):553–563CrossRefGoogle Scholar
  40. Zaheer M, Reddy VD, Giri CC (2016) Enhanced daidzin production from jasmonic and acetyl salicylic acid elicited hairy root cultures of Psoralea corylifolia L.(Fabaceae). Nat Prod Res 30(13):1542–1547CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  1. 1.Laboratory for Plant Physiology, Department of Biology, Faculty of ScienceUniversity of SarajevoSarajevoBosnia and Herzegovina
  2. 2.Laboratory for Research and Protection of Endemic Resources, Department of Biology, Faculty of ScienceUniversity of SarajevoSarajevoBosnia and Herzegovina
  3. 3.Central Laboratories and Research Support Faculty of Science, Centre of the Region Haná for Biotechnological and Agricultural ResearchPalacky UniversityOlomoucCzech Republic
  4. 4.Department of Genetic Resources for Vegetables, Medicinal and Special Plants, Centre of the Region Haná for Biotechnological and Agricultural ResearchCrop Research InstituteOlomoucCzech Republic

Personalised recommendations