Page 109 - Book of Abstracts

5 International Scientific Online Conference DOI: https://doi.org/10.15414/2021.9788055224015

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DEVELOPMENT OF BIOTECHNOLOGICAL METHOD OF DELPHINIUM ELATUM L.
CULTIVATION
Romana Petrina, Diana Zahorodnia, Kateryna Kniazieva, Olha Shved, Vira Lubenets
Lviv Polytechnic National University, Lviv, Ukraine; E-mail.: romanna.o.petrina@lpnu.ua

Species that belong to the family Delphinium are interesting representatives of the plant
kingdom that have a practical use. This is caused by the fact that they contain diterpene
alkaloids, which show analgesic, anti-inflammatory, antimicrobial, anticancer, antiarrhythmic
activity. Also, they contain phenolic compounds, which show antibacterial, antifungal, immune-
stimulating, analgesic, antioxidant activity. Delphinium elatum L. contains diterpene alkaloids,
that show cytotoxic activity against the cell lines of nasopharynx cancer, they are resistant to
cell lines of lungs, prostate, and nasopharynx cancer. Information about D. elatum, which grows
in Ukrainian Carpathians and is included in the Ukrainian Red Book, is limited, there are no
publications about its composition and application.
The plant tissue culture method has advantages over classical harvesting of medicinal
plants and growing plants in the field. The use of in vitro technology allows to regulate plant
cells' growing and secondary metabolites accumulation by optimizing culture medium contain
and cultivation conditions. Callus biomass produces an infinite amount and can be used as a raw
material for medical purposes because it is ecologically clean. Moreover, it is not contaminated
by chemicals, pesticides and herbicides.
We used for our experiment D. elatum seeds that had previously been soaked in water for
24 hours. The seeds were sterilized using 30 % hydrogen peroxide solution and 70% ethanol
solution. Then it was rinsed three times with sterile distilled water. The seeds were introduced
into the culture medium in vitro accommodation and sterile plants were obtained.
Leaves and stems of the plants were used as explants for callus genesis induction in the
Murashige-Skuga culture medium with adding of phytohormones. 0.1–3.0 mg/l auxin – indole
acetic acid (IAA), α-naphthyl-1-acetic acid (NAA) in various combination and 0.5 mg/l 6-
aminopurine (kinetin) were used as phytohormones. The culture medium also contained 8–9
g/l agar, рНwas 5.7. Cultivation was performed with a photoperiod of 16/8 hours (light/dark),
illumination 2000 Lux, temperature 26 ±2 °C, relative humidity 60–70 % for 50 days (2
passages). All experiments were performed in 3 replicates and the results were statistically
processed. The influence of phytohormones and explant type on culture growth rate has been
studied. The highest intensity of callus genesis was found in the case of leaf explants use and the
culture medium with 1.0 mg/l NAA and 2.0 mg/l IAA as phytohormones.
All explants formed a callus. The obtained biomass was light yellow and had fluffy
consistency. The best percentage of callus genesis was observed for leaves explants (84 %),
then for steams explants (72 %). The frequency of callus genesis was determined as the ratio
of the explants with callus number to the total explants number in percent every 3 weeks of
cultivation.
The cultivation conditions in vitro of D. elatum were selected. The influence of
concentration and phytohormones contain in culture medium on callus biomass growth rate
was studied. The biotechnological method of biomass production was developed to preserve
plant biodiversity. The ecologically clean raw material was obtained to identify its secondary
metabolites.
Keywords: Delphinium elatum, cultivation, biotechnological method, callus genesis, explants,
phytohormones.

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5 International Scientific Conference Agrobiodiversity for Improving the Nutrition, Health, Quality of Life and |108
Spiritual Human Development
November 3 2021
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