Page 101 - Book of Abstracts

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

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AMINO ACIDS ACCUMULATION IN CICHORIUM INTYBUS L. "HAIRY" ROOTS
Nadiia Matvieieva , Hanna Shutava , Ljudmila Tychinskaya , Eugenij Skakovsky
3
2
3
1
1 Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences
of Ukraine, Kyiv, Ukraine; Email: joyna56@gmail.com
2 Central Botanical Garden of the National Academy of Sciences of Belarus, Minsk, Belarus
Institute of Physical Organic Chemistry of the National Academy of Sciences of Belarus,
3
Minsk, Belarus
Agrobacterium rhizogenes-mediated transformation is a well-known method for plant
genetic transformation. After plant parts (leaves, stems, cotyledons, etc.) cocultivation with
soil bacteria "hairy" roots growth can be initiated. These roots are of practical interest
because of the synthesis of different compounds including the chemicals which can be used
in medicine. Polyphenols, sugars, amino acids, essential oils can be synthesized in the cells
of “hairy” roots. The level of the accumulation of the compounds in root clones can differ
because of the effect of the site of incorporation of bacterial genes (rol genes) in the plant
genome (independent transformational events).
In the case of "hairy" roots using for the creation of pharmaceuticals, it is important to
maximize the identification of all groups of biologically active compounds that can be
extracted during extraction. These metabolites include amino acids, which have a number
of valuable pharmaceutical properties. However, there is no information about their
accumulation in chicory "hairy" roots. In this work, we compared the content of amino acids
in two "hairy" root clones cultivated in vitro.
"Hairy" roots of Cichorium intybus L. were grown under standard conditions on
solidified Murashige and Skoog nutrient medium supplemented with 20g/L sucrose,
collected, lyophilized, and crushed. The content of amino acids was studied by Nuclear
magnetic resonance spectroscopy. To prepare the extracts, 0.07 g of each of the samples was
added to 10 ml glass containers and 2 ml of deuterated water (D2O, degree of deuteration – 99.8
%) was added. Samples in closed containers were kept at room temperature with periodic
stirring for 24 hours. Then the extracts were filtered and made into ampoules for the
subsequent recording of NMR spectra. The spectra on 1H and 13C nuclei were recorded on an
AVANCE-500 NMR spectrometer (Bruker, Germany) with operating frequencies of 500 and 125
MHz for 1H and 13C nuclei, respectively, at a temperature of 293 K.
Differences in the content of the amino acid of root clones No1 and No2 were found (5.83
and 8.84 %, respectively). Glutamine, Asparagine, Alanine and Arginine HCl and Threonine
content was greater in the root clone No2 compared clone No1 (1.59 and 0.55 %; 2.68 and 1.78
%; 0.2 and 0.08 %; 3.45 and 2.72 %; 0.21 and 0.09 %, respectively). At the same time, there
were no differences in the content of Isoleucine, Valine, Leucine, Lysine didn’t differ in studied
root clones.
So, differences in the content of the amino acids were found in two Chicory “hairy” root
lines. These differences can be explained by the effect of the position of bacterial genes
transferred in the plant genome after the Agrobacterium-mediated transformation.
Keywords: Cichorium intybus, "hairy" roots, Agrobacterium rhizogenes-mediated transformation,
amino acids.
Acknowledgments
The work was partially supported by a grant № II-2-21 of the National Academy of Sciences of Ukraine
and № B21-UKRG-005 of BRFFR.

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