Page 136 - Zbornik_Konf_2019

https://doi.org/10.15414/2019.9788055220703

4 International Scientific Conference Abstracts Book
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TRANSGENESIS AS A TOOL FOR WIDENING OF THE GENETIC VARIATIONS AND USING OF
MEDICINAL PLANTS
Yana Sindarovska, Nikolay Kuchuk

1 The Institute of Cell Biology and Genetic Engineering, NAS of Ukraine, Kyiv, Ukraine;
E-mail.: sindarovskaya@ukr.net
Medicinal plants are the richest bioresources of drugs for traditional and modern
medicine. Their compounds are included in herbal health care formulations, herbal nutrients,
and herbal-based cosmetics. To obtain plants with new characteristics and properties the
transgenic technology can be used. Agrobacterium rhizogenes is a soil bacterium and a natural
tool for transgenesis: it causes hairy root formation at the site of infection. Hairy roots can
produce higher levels of useful secondary metabolites, change their composition, or reveal
novel bioactive compounds valuable for herbal medicine. Plants regenerated from hairy root
culture often have an altered phenotype.
The aim of our work is obtaining transgenic medicinal plants via hairy root cultures and
estimation of their characteristics and properties.
For obtaining transgenic plants we chose five medicinal herbs often used in Ukrainian
folk medicine and pharmaceutical industry, namely Silybum marianum (L.) Gaertn., Matricaria
chamomilla L., Origanum vulgare L., Lavandula angustifolia Mill., and Lophanthus anisatus
(Nutt.) Benth. Seeds were surface-sterilized for obtaining in vitro aseptic plant cultures. Two
sterilizing methods were tested: rinsing in 75 % ethanol following 4 % hypochloride solution
or rinsing in 70 % ethanol with 0.1 % Triton-X100, and next washing in distilled water. Then
seeds were placed on nutrient Murashige-Skoog (MS) agar medium. Seedling was germinated
and grew in the growth chamber at 24 C. Adult plants were cloned, and the regeneration
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media were selected. Species showed well-developed regeneration protocol were
transformed by A. rhizogenes to obtain hairy root culture.
Testing of seed sterilization methods demonstrated that using of hypochloride solution
is more effective against fungal contamination but affect essentially on germination efficiency.
The most effective protocol was developed for L. anisatus where seed germination was about
100 % with no contamination. L. angustifolia and M. chamomilla had less viable seeds after
sterilization but germinated seedlings grew well and produced stable in vitro culture. Used
sterilization protocols were not effective for S. marianum and O. vulgare species, and although
a few in vitro adult plants were obtained for these species, they finally died because of severe
fungi attack. Species were also checked for cloning and regeneration efficiency. L. anisatus, L.
angustifolia, O. vulgare, and S. marianum showed effective growth and root formation on MS
medium after shoot/stem cutting. M. chamomilla has leaves arranged in rosettes and
sometimes two rosettes appear so it was possible to divide them for cloning; root formation
and plant growth were good on MS medium. Regeneration potential was checked for L.
anisatus, L. angustifolia, and M. chamomilla. New plants were nicely regenerated on MS
medium supplemented with phytohormones 1 mg/L BAP and 0.1 mg/L IAA from L. anisatus
and L. angustifolia explants. Preliminary results showed that nutrient media contained kinetin
will be more suitable for M. chamomilla regeneration. A. rhizogenes‐mediated transformation
of L. anisatus and L. angustifolia is in the progress.
The results demonstrate that some species are very convenient for in vitro
manipulations and can be used for obtaining medicinal transgenic plants with altered
properties.

Keywords: medicinal plants, Agrobacterium rhizogenes, hairy roots, transgenic plants.

|136 4 International Scientific Conference Agrobiodiversity Nutrition, Health and Quality of Human and Bees Life
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September 11–13, 2019