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Effect of the genetic transformation on the antioxidant activity of Althaea officinalis L., Artemisia vulgaris L. and Artemisia tilesii Ledeb. «hairy» roots
SUMMARY. Genetic transformation using Agrobacterium rhizogenes is a well-known method for induction of «hairy» root cultures of different plant species. During the process of transformation bacterial rol genes are incorporated into the genome of the forming roots. Incorporation of rol genes, known as stimulators of plant secondary metabolism, as well as contact with phytopathogenic bacteria, can affect a number of growth parameters of «hairy» roots, in particular, mass increase, accumulation of secondary metabolites, the activity of ferments etc. The study of changes initiated by Agrobacterium-mediated transformation after long-term cultivation of transgenic plant material has a special interest. In this work antioxidant potential of Althaea officinalis L., Artemisia tilesii Ledeb., Artemisia vulgaris L. «hairy» roots obtained using Agrobacterium rhizogenes mediated transformation (A4 strain carrying human interferon-α2b gene) was studied. Some parameters such as the activity of antioxidant enzymes (catalase and superoxide dismutase), total content of flavonoids, antioxidant and reducing activity of «hairy» root extracts were evaluated and compared with the same parameters in the roots of in vitro cultivated plants. The presence of foreign genes in studied «hairy» roots after long-term cultivation was confirmed by PCR analyses. Significant variability of catalase and SOD activity in different lines of «hairy» roots as well as the difference between these parameters and the control one were revealed. In particular, we found root lines that were characterized by increased activity of both enzymes (4.4 times increased catalase activity and twice the activity of SOD compared to the activity of extracts from the roots of control plants). The possibility of a significant increase in the content of total flavonoids (up to 4.6 times compared to the control) and an increase in the levels of antioxidant and reducing activities in some samples of transgenic roots was demonstrated. The maximum content of flavonoids in the «hairy» roots of A. officinalis, A. vulgaris and A. tilesii was 4.60 ± 0.19, 4.55 ± 0.36 and 9.21 ± 1.28 mg/g of fresh weight, respectively. Thus, it was found that genetic transformation led to changes in the synthesis of metabolites with antioxidant properties and enzyme activity of the antioxidant defense system, and such changes were observed 5–8 years after the induction of «hairy» roots. These results indicate a prolonged effect of genetic transformation on the functioning of plant cells of different species, in particular, marshmallow, and two wormwood species. The detected effect of increasing the content of flavonoids and level of antioxidant activity in most samples cultivated over a long period of time can be used to obtain «hairy» roots – producers of compounds with antioxidant properties. Key words: «hairy» roots, antioxidant activity, flavonoids, catalase, superoxide dismutase, Althaea officinalis, Artemisia tilesii, Artemisia vulgaris
Tsitologiya i Genetika 2021, vol. 55, no. 6, pp. 40-50
E-mail: bogdanovych_tais
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