The production of plant biomass by alternative biotechnological method is economically advantageous and promising as it contains secondary metabolites, which is a plant source, is environmentally friendly, is obtained throughout the year, regardless of season and weather conditions. This is especially important for rare and endangered plants, which are a valuable raw material base for pharmacy and medicine.
The cultivation in this way of plants that have not yet been investigated and the detection of biologically active substances in their callus biomass is an economically and environmentally beneficial alternative to modern sources of these compounds.
The aim of the work is to obtain the Adonis vernalis and Aquilegia nigricans callus biomass using the biotechnological method of culture of cells, tissues and organs in vitro and study it on the content of secondary metabolites such as cardiac glycosides, common phenols and alkaloids.
Adonis vernalis and Aquilegia nigricans are plants of the Ranunculaceae family, listed in the Red Book of Ukraine. Adonis vernalis is used in medical practice for the preparation of cardiological agents, contains glycosides (cymarin, adoniside, adenyvirnozid, adonitoksin), triterpene saponins, flavonoids. Aquilegia nigricans have been little studied, in the literature very scanty information about it, on the chemical composition, properties. The literature describes Aquilegia vernalis, which contains alkaloids, cyanogen compounds and ascorbic acid.
Adonis vernalis and Aquilegia nigricans were obtained on the Murashige-Skoog (MC) medium with the addition of growth regulators (indolylocytic acid (IOC), naphthylactoacid (NOC), kinetin) and vitamins (B1, B2, B6, folic acid, biotin, nicotinamide , pantothenate).
Adonis vernalis caluse biomass was obtained on the MS medium with the addition of IOC, NOC and kinetin in the amount of 2.0 mg / l, 1.0 mg / l, 0.1 mg / l respectively at 23 ° C, illuminating 2000 lux, photoperiod 16/8. Different conditions (temperature, lighting), type of explant and different concentrations of phytohormones have been experimentally selected for the production of Aquilegia nigricans for calamus biomass. The best growth characteristics were observed at 23 ° C on light in an MS medium containing IOC - 0.2 mg / L, NOC - 0.2 mg / L, kinetine - 0.1 mg / L; with an IOC content of 0.3 mg / L, a NOC of 1.0 mg / L, a kinetine of 0.1 mg / L and an IOC of 0.3 mg / L, a NOC of 0.3 mg / L, kinetin - 0.5 mg / L.
Extracts of calamus biomass and vegetative raw materials of Adonis vernalis and Akilegia nigricans in 70% ethanol were obtained. Identification of common phenols, cardiac glycosides and alkaloids with the help of color specific reactions has been performed. Quantitative determinations of the presence of BAR in extracts were also carried out using a spectrophotometer. The extracts of the Adonis vernalis and Aquilegia nigricans were found to contain less common phenols and more cardiac glycosides than in intact plants.
Key words: Adonis vernalis, Aquilegia nigricans, cultivation, growth regulators, kalus, cardiac glycosides, general phenols.
1. Butenko R. H. Kultura klityn roslyn i biotekhnolohiia / Butenko R. H. – M.: Nauka, 1986. – 286 s.
2. Chekman I. S. Eksperymentalne y klinichne vyvchennia sertsevykh hlikozydiv / I. S.Chekman // Praktykuiuchyi likar.– 2014. – № 3.– S. 54–59.
3. Vasylev Y. S. Steroydnыe hlykozydы yz kulturы kletok dyoskorey, ykh metabolyzm y byolohycheskaia aktyvnost / Y. S. Vasylev, V. A. Paseshnychenko // Uspekhy byol. khymyy. – 2000. –T. 40. – S. 153–204.
4. Karpov P. A. Vozmozhnosty yspolzovanyia kulturы tkanei Yucca macrocara dlia poluchenyia steroydnыkh hlykozydov / P. A. Karpov // Dopovidi AN Ukrainy. – 2000. – № 9. – S. 180–85.
5. Kunakh V. A. Biotekhnolohiia likarskykh roslyn. Henetychni ta fizioloho-biokhimichni osnovy / V. A. Kunakh. – K. : Lohos, 2005. – 730 s.
6. Hershtun A. Kultyvuvannia horytsvitu vesnianoho (Adonis vernalis) v umovakh in vitro / A. Hershtun, R. Petrina // Visnyk Nats. un-tu “Lvivska politekhnika”. Seriia “Khimiia, tekhnolohiia rechovyn tazastosuvannia”. – 2016. – № 841. – S.133–137.
7. Gafta D., Muncaciu S., Csergo A. Morphometric va71 riation in a rare endemic Aquilegia (Ranunculaceae) in the Carpathians // Plant Biosystems. – Vol. 140. – No. 3. – 2006. – P. 297–306.
8. Assyov, B. and Petrova, A. (Eds.). 2012. Conspectus of the Bulgarian Vascular Flora: Distribution maps and floristic element. Bulgarian Biodiversity Foundation, Sofia.
9. Oprea, A. and Sirbu, C. 2013. The Vascular Flora of Rarau Massif (Eastern Carpathians, Romania). Note II. Memoirs of the Scientific Sections of the Romanian Academy XXXVI: 17–36.
10. Coldea, G., Stoica, I.-A., Puşcaş M, Ursu, T, Oprea, A, Intrabiodiv Consortium. 2009. Alpine–subalpine species richness of the Romanian Carpathians and the current conservation status of rare species. Biodiversity Conservation. 18. P. 1441–1458.
11. Likarski roslyny: entsyklop. dovidnyk / A. M. Hrodzinskyi. – K.: Vydavnytstvo “Ukrainska Entsyklopediia” im. M. P. Bazhana, Ukrainskyi vyrobnycho-komertsiinyi tsentr “Olimp”, 1992.– 544 s.
12. Oksman-Caldentey K. M. and Inzé D. Plant cell factories in the post genomic era: new ways to produce designer secondary metabolites. Trends Plant Sci, 2004, P. 433–440.
13. Rao, S. Ramachandra and Ravishankar G. A. Plant cell cultures: Chemical factories of secondary metabolites. Biotechnology Advances, 2002. 20, P. 101–153.
14. Parr A. J. The Production of Secondary Metabolites by Plant cell cultures. Journal of Biotechnology, 1989. 10, P. 1–26.
15. Solich P., Sedliakova V., Karlicek R. Spectrophotometric determination of cardiac glycosides by flow-injection analysis. Anal Chim Acta. 1992; 269(2). P. 199–203.
16. Waterman P. G., Mole S. Analysis of Phenolic Plant Metabolites. Blackwell Scientific Publications. London. 1994.