FORMATION AND IVESTIGATION OF CALLUS MASS OF DELPHINIUM ELATUM L.

Authors

DOI:

https://doi.org/10.11603/2312-0967.2020.2.11205

Keywords:

Delphinium elatum L., growth regulators, callus, phenolic compounds, flavonoids, extract, antimicrobial activity, antioxidant activity

Abstract

The aim of the work. Introduce to culture in vitro Delphinium elatum L., investigate the effect of phytohormones on plant cell growth processes, determine optimal conditions of cultivation in vitro. Obtain extracts of biomass and plant materials of D. elatum, conduct a comparative study of biologically active compounds, antimicrobial and antioxidant properties.

Materials and Methods. Cultivation of D. elatum was performed on a modified Murashige and Skoog culture medium. Extracts were obtained by maceration method, as extractant used ethanol solutions in concentrations of 20, 40, 70 and 90 %. The total phenolic content was determined by spectrophotometric method using Folin-Ciocalteu reagent, the total flavonoid content – by colorimetric method with aluminium chloride. Antimicrobial activity was investigated on standard and clinical strains of microorganisms by diffusion in agar, antioxidant activity – by spectrophotometric method by interaction with stable chromogen radical 2,2- diphenyl-1-picrylhydrazyl.

Results and Discussion. The optimal stratification scheme of D. elatum seeds with the highest yield of aseptic explants was selected: consecutive mechanical treatment, wetting with cold sterile water, refrigerator ageing, treatment with 70% ethanol, sodium hypochlorite and sterilized bidistilled water.

The composition of the Murashige and Skoog culture medium, supplemented with phytohormones BAP and 2,4-D, where the highest specific growth rates and the shortest time of doubling the biomass of calluses were observed, was determined. The callus mass was obtained after 35 days of cultivation.

The total phenolic and flavonoids contents of D. elatum extracts were studied. It was determined that extracts of both callus biomass and plant material obtained with 40 % ethanol solution contain the highest number of phenolic compounds and flavonoids.

Antimicrobial and antifungal activity of D. elatum extracts in relation to standard and clinical strains of some microorganisms was investigated. It was determined that maximum activity was shown by D. elatum extracts obtained with 40 % and 70 % ethanol solutions.

Conslusions. As a result of the research it was established that the D. elatum callus biomass in terms of content of phenolic compounds, flavonoids and pharmacological activity is not inferior to vegetable raw materials and can be used as an equivalent medicinal raw material.

Author Biographies

O. S. Khropot, Lviv Polytechnic National University

PhD student, Department of Technology of Biologically Active Substances, Pharmacy and Biotechnology

Ye. V. Bazavluk, Lviv Polytechnic National University

student, Department of Technology of Biologically Active Substances, Pharmacy and Biotechnology

R. T. Konechna, Lviv Polytechnic National University

PhD (Pharmacy), Associate Professor of the Department of Technology of Biologically Active Substances, Pharmacy and Biotechnology

I. I. Hubytska, Lviv Polytechnic National University

PhD (Сhemistry), Associate Professor of the Department of Technology of Biologically Active Substances, Pharmacy and Biotechnology

Y. Т. Konechnyi, Danylo Halytsky Lviv National Medical University

PhD student, Department of Microbiology

I. Jasicka-Misiak , University of Opole, Poland

doctor habilitatus, Professor at the Department of Analytical and Ecological Chemistry

P. P. Wieczorek, University of Opole, Poland

doctor habilitatus, Professor, Dean of the Faculty of Chemistry

V. P. Novikov, Lviv Polytechnic National University

DS (Сhemistry), Professor, Head of the Technology of Biologically Active Substances, Pharmacy and Biotechnology Department

References

Didukh YaP. Red Data Book of Ukraine. Flora [Червона книга України. Рослинний світ] Kyiv: Hlobalkonsaltynh; 2009. Ukrainian.

Konechna RT, Konechnyi YuT, Petrina RO, Shykula RH, Wieczorek P, Jasicka-Misiak I, et al. Obtaining and research of callus mass of Gentiana lutea L. roots. Res J Pharm Biol. Chem. Sci. 2015;6(4): 1490-5.

Shimizu K, Yogai T, Hashimoto F, Sakata Y. Plant regeneration from suspension cell culture of Delphinium Hort J. 2004;73(5): 435-40. Available from: https://doi.org/10.2503/jjshs.73.435 DOI: https://doi.org/10.2503/jjshs.73.435

Hosokawa K, Koiwa H, Yamamura S. In vitro adventitious shoot formation on petioles of commercial cultivars of Delphinium. Sci Hortic. 2001;90(1-2): 143-50. Available from: https://doi.org/10.1016/S0304-4238(00)00249-1 DOI: https://doi.org/10.1016/S0304-4238(00)00249-1

Murphy KP, Santamaria JM, Davibs WJ, Lumsden PJ. Ventilation of culture vessels. I. Increased ’growth in vitro and survival ex vitro of Delphinium. J. Hortic Sci Biotechnol. 1998;73: 725-9. Available from: https://doi.org/10.1080/14620316.1998.11511039 DOI: https://doi.org/10.1080/14620316.1998.11511039

Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. plant. 1962;15(3): 473-497. Available from: https://doi.org/10.1111/j.1399-3054.1962.tb08052.x DOI: https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

Konechna RT, Petrina RO, Novikov VP, Konechnyi YuT, Shykula RH, Korniichuk OP. [Research of callus mass extracts Carlina acaulis L.]. Ukr Bioph J. 2015;4(39): 57-61.

Kokotkiewicz A, Bucinski A, Luczkiewicz M. Light and temperature conditions affect bioflavonoid accumulation in callus cultures of Cyclopia subternata Vogel (honeybush). PCTOC. 2014;118(3): 589-93. Available from: http://dx.doi.org/10.15294/biosaintifika.v12i1.21704 DOI: https://doi.org/10.15294/biosaintifika.v12i1.21704

State Pharmacopoeia of Ukraine. 1st edition. Appendix 1. [Державна Фармакопея України] Kharkiv: State Enterprise "Scientific Expert pharmacopoeia center"; 2004. Ukrainian.

Wilczyńska A. Phenolic content and antioxidant activity of different types of Polish honey – A short report. Pol J Food Nutr Sci. 2010;6: 309-13.

Meda A, Lamien Ch, Romito M, Millogo J, Nacoulma O. Determination of the total phenolic, flavonoid and proline contents in Burkina Fasan honey, as well as their radical scavenging activity. Food Chem. 2005;91: 571-7. Available from: https://doi.org/10.1016/j.foodchem.2004.10.006 DOI: https://doi.org/10.1016/j.foodchem.2004.10.006

Downloads

Published

2020-06-23

How to Cite

Khropot, O. S., Bazavluk, Y. V., Konechna, R. T., Hubytska, I. I., Konechnyi Y. Т., Jasicka-Misiak , I., … Novikov, V. P. (2020). FORMATION AND IVESTIGATION OF CALLUS MASS OF DELPHINIUM ELATUM L. Pharmaceutical Review Farmacevtičnij časopis, (2), 5–15. https://doi.org/10.11603/2312-0967.2020.2.11205

Issue

No. 2 (2020)

Section

Phytochemical researches

License

The authors who publish their materials in this journal agree with the following terms:

  • Authors reserve the right to authorship of their work and assign to the journal the right to first publish this work under the terms of the Creative Commons Attribution License, which allows other persons to freely distribute the published work with a mandatory reference to the authors of original work and the first publication of work in this journal Результат пошуку зображень за запитом "CC-BY logo".
  • Authors have the right to make independent extra-exclusive work agreements in the form they are published by this journal (for example, posting work in an electronic repository of an institution or publishing as part of a monograph), provided that the link to the first publication of the work in this journal is maintained.

Journal policy allows and encourages publication of manuscripts on the Internet (for example, in institutions repositories or on personal websites), both before the publication of this manuscript and during its editorial review, as it contributes to productive scientific discussion and positively affects the efficiency and dynamics of the citation of the published work (see The Effect of Open Access).