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ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)

 

Biotechnologia Acta  V. 15, No. 3, 2022
P. 13-22. Bibliography 26, Engl.
UDC: 604.6:577.13
https://doi.org/10.15407/biotech15.03.013

PECULIARITIES OF THE GROWTH OF  Artemisia tilesii Ledeb. “HAIRY” ROOTS WITH DIFFERENT FOREIGN GENES

Т. А. Bohdanovych 1, B. V. Morgun 1, 2, О. R. Lakhneko 1, 2, А. М. Shakhovsky 1, N. А. Matvieieva 1

1Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, Kyiv
2 National Technical University of Ukraine “Igor Sikorsky Kyiv Polytechnic Institute”

Aim. To compare Artemisia tilesii “hairy” root lines with different transferred genes in terms of the relationship between the total content of flavonoids, the levels of antioxidant activity (AOA) and reducing power (RP), as well as the activity of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), rolB and rolC genes.

Methods. We compared the root lines Nos. 10 and 16, obtained by transformation with the wild Agrobacterium rhizogenes strain A4, lines Nos. 2 and 4, obtained using A. rhizogenes carrying pSV124 vector with ifn-α2b and nptII genes, as well as the roots of non-transformed plants that were cultivated in vitro. The presence and activity of rolB, rolC, PAL, and CHS genes were determined by PCR and real-time PCR, respectively. The content of flavonoids, AOA and RP were determined by standard tests with AlCl3, DPPH (2,2-diphenyl-1-picrylhydrazyl) and K3[Fe(CN)6] accordingly.

Results. The content of flavonoids in most of the lines was higher than in the control, and correlated with AOA and RP. Roots No. 10 were characterized by the fastest growth, which coincided with higher activity of rolB and rolC genes. The activities of PAL and CHS in “hairy” roots were lower than those in non-transformed ones.

Conclusions. Root lines carrying only rolB and rolC and lines with additional ifn-α2b and nptII genes had similar ranges of flavonoids concentration, AOA and RP levels that exceeded those in the control. The dependence of the root growth rate, and lack of the dependence of the flavonoids content with the activity of the rol genes were demonstrated. PAL activity inversely correlated with flavonoids content in all experimental lines, which may be the result of overproduction of compounds in transgenic roots.

Key words: Artemisia tilesii Ledeb., Agrobacterium rhizogenes, “hairy” roots, flavonoids, rol genes, PAL and CHS genes, real-time PCR, antioxidant activity.

© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2022