Skip to main content
SpringerLink
Log in

Effects of Genetic Transformation on the Antioxidant Activity of “Hairy” Roots of Althaea officinalis L., Artemisia vulgaris L., and Artemisia tilesii Ledeb.

  • Published:
Cytology and Genetics Aims and scope Submit manuscript

Abstract

Agrobacterium rhizogenes-mediated genetic transformation is a well-known method for the induction of “hairy” root cultures of different plant species that allows to incorporate the bacterial rol genes into the plant genome. Introduction of the rol genes known as the stimulators of plant secondary metabolism, as well as a contact with phytopathogenic bacteria, may affect different growth parameters, such as weight increase, accumulation of secondary metabolites, and activity of enzymes, etc. after a long-term cultivation study of changes induced by Agrobacterium-mediated transformation and a transfer of foreign genes into the plant genome is of special interest. The activity of the enzymes (catalase and superoxide dismutase) belonging to the plant antioxidant defense system, as well as the total flavonoid content, the antioxidant and reducing activity of extracts from “hairy” root cultures, including Althaea officinalis, Artemisia vulgaris, and Artemisia tilesii, which were earlier obtained through the A. rhizogenes-mediated transformation (A4 strain with the human interferon ifn-α2b gene) were compared. PCR analysis confirmed the presence of the transferred genes in the corresponding “hairy” roots after long-term cultivation. A significant variability has been observed in the catalase and superoxide dismutase (SOD) activities among different “hairy” root lines. In particular, some lines characterized by increased activity of both enzymes have been identified. Catalase and SOD activities increased, respectively, 4.4 times and twice compared with the activity of extracts from the roots of the control plants. The possibility of significant increases in the total flavonoid content (by 4.6 times) and the levels of antioxidant and reducing activities in some samples of transgenic roots have been shown. The maximum flavonoid contents reached in the “hairy” roots of A. officinalis, A. vulgaris, and A. tilesii 4.60 ± 0.19, 4.55 ± 0.36, and 9.21 ± 1.28 mg/g of fresh weight. Thus, it has been demonstrated that genetic transformation induces changes in both the synthesis of metabolites with antioxidant properties and the activities of the antioxidant defense system enzymes. Changes were retained during 5–8 years after the induction of “hairy” roots. These results confirm a prolonged effect of genetic transformation on the functions of cells in the medicinal plant species, including marshmallow and two wormwood species. The established effect of increasing the flavonoid content and the level of antioxidant activity in the majority of samples after a long-term cultivation may be used for obtaining “hairy” roots and their usage as producers of compounds with antioxidant properties.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.
Fig. 5.
Fig. 6.

Similar content being viewed by others

REFERENCES

  1. Bulgakov, V.P., Gorpenchenko, T.Y., Veremeichik, G.N., et al., The rolB gene suppresses reactive oxygen species in transformed plant cells through the sustained activation of antioxidant defense, Plant Physiol., 2012, vol. 158, pp. 1371–1381. https://doi.org/10.U04/pp.1U.191494

  2. El-Esawi, M.A., Elkelish, A., Elansary, H.O., et al., Genetic transformation and hairy root induction enhance the antioxidant potential of Lactuca serriola L., Oxid. Med. Cell Longev., 2017, vol. 2017, art. 5604746. https://doi.org/10.1155/2017/5604746

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Gabr, A.M.M., Sytar, O., Ghareeb, H., et al., Accumulation of amino acids and flavonoids in hairy root cultures of common buckwheat (Fagopyrum esculentum), Physiol. Mol. Biol. Plants, 2019, vol. 25, pp. 787–797. https://doi.org/10.1007/s12298-019-00669-1

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Gharari, Z., Bagheri, K., Danafar, H., et al., Enhanced flavonoid production in hairy root cultures of Scutellaria bornmuelleri by elicitor induced over expression of MYB7 and FNS2 genes, Plant Physiol. Biochem., 2020, vol. 148, pp. 35–44. https://doi.org/10.1016/j.plaphy.2020.01.002

    Article  CAS  PubMed  Google Scholar 

  5. Giri, A. and Narasu, M.L., Transgenic hairy roots: recent trends and applications, Biotechnol. Adv., 2000, vol. 18, pp. 1–22. https://doi.org/10.1016/S0734-9750(99)00016-6

    Article  CAS  PubMed  Google Scholar 

  6. Khezerluo, M., Hosseini, B., and Amiri, J., Sodium nitroprusside stimulated production of tropane alkaloids and antioxidant enzymes activity in hairy root culture of Hyoscyamus reticulatus L., Acta Biol. Hung., 2018, vol. 69, pp. 437–448. https://doi.org/10.1556/018.69.2018.4.6

    Article  CAS  PubMed  Google Scholar 

  7. Kim, S.-E., Lee, C.-J., Ji, C., et al., Transgenic sweet potato plants overexpressing tocopherol cyclase display enhanced a-tocopherol content and abiotic stress tolerance, Plant Physiol. Biochem., 2019, vol. 144, art. 436444. https://doi.org/10.1016/j.plaphy.2019.09.046

    Article  CAS  Google Scholar 

  8. Kohsari, S., Rezayian, M., Niknam, V., et al., Antioxidative enzymes activities and accumulation of steroids in hairy roots of Trigonella, Physiol. Mol. Biol. Plants, 2020, vol. 26, pp. 281–288. https://doi.org/10.1007/s12298-019-00753-6

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Kumar, S. and Pandey, A.K., Chemistry and biological activities of flavonoids: an overview, Sci. World J., 2013, 2013, art. 162750. https://doi.org/10.1155/2013/162750

    Article  CAS  Google Scholar 

  10. Matvieieva, N.A., Shakhovsky, A.M., Belokurova, V.B., et al., Artemisia tilesii Ledeb hairy roots establishment using Agrobacterium rhizogenes-mediated transformation, Prep. Biochem. Biotechnol., 2015, vol. 46, pp. 342–345 https://doi.org/10.1080/10826068.2015.1031393

    Article  CAS  Google Scholar 

  11. Matvieieva, N., Drobot, K., Duplij, V., et al., Flavonoid content and antioxidant activity of Artemisia vulgaris L. “hairy” roots, Prep. Biochem. Biotechnol., 2019, vol. 49, pp. 82–87. https://doi.org/10.1080/10826068.2018.1536994

    Article  CAS  PubMed  Google Scholar 

  12. Matvieieva, N., Morgun, B., Lakhneko, O., et al., Agrobacterium rhizogenes-mediated transformation enhances the antioxidant potential of Artemisia tilesii Ledeb., Plant Physiol. Biochem., 2020, vol. 152, pp. 177–183. https://doi.org/10.1016/j.plaphy.2020.04.020

    Article  CAS  PubMed  Google Scholar 

  13. Muthusamy, B. and Shanmugam, G., Analysis of flavonoid content, antioxidant, antimicrobial and antibiofilm activity of in vitro hairy root extract of radish (Raphanus sativus L.), Plant Cell Tiss. Organ Cult., 2020, vol. 140, pp. 619–633.https://doi.org/10.1007/s11240-019-01757-6

    Article  CAS  Google Scholar 

  14. Pekal, A. and Pyrzynska, K., Evaluation of aluminium complexation reaction for flavonoid content assay, Food Anal. Methods, 2014, vol. 7, pp. 1776–1782. https://doi.org/10.1007/s12161-014-9814-x

    Article  Google Scholar 

  15. Pietta, P.G., Flavonoids as antioxidants, J. Nat. Prod., 2000, vol. 63, pp. 1035–1042. https://doi.org/10.1021/np9904509

    Article  CAS  PubMed  Google Scholar 

  16. Reis, A., Boutet-Mercey, S., Massot, S., et al., Isoflavone production in hairy root cultures and plantlets of Trifolium pretense, Biotechnol. Lett., 2019, no. 3, pp. 427–442. https://doi.org/10.1007/s10529-018-02640-8

  17. Sahayarayan, J., Udayakumar, R., Arun, M., et al., Effect of different Agrobacterium rhizogenes strains for in-vitro hairy root induction, total phenolic, flavonoids contents, antibacterial and antioxidant activity of Cucumis anguria L., Saudi J. Biol. Sci., 2020, vol. 27, pp. 2972–2979. https://doi.org/10.1016/j.sjbs.2020.08.050

    Article  CAS  Google Scholar 

  18. Singh, H., Dixit, S., Verma, P.C., et al., Evaluation of total phenolic compounds and insecticidal and antioxidant activities of tomato hairy root extract, J. Agric. Food Chem., 2014, vol. 62, pp. 2588–2594. https://doi.org/10.1021/jf405695y

    Article  CAS  PubMed  Google Scholar 

  19. Tavassoli, P. and Safipour Afshar, A., Influence of different Agrobacterium rhizogenes strains on hairy root induction and analysis of phenolic and flavonoid compounds in marshmallow (Althaea officinalis L.), 3 Biotech, 2018, vol. 8, p. 351. https://doi.org/10.1007/s13205-018-1375-z

  20. Thwe, A., Arasu, M.V., Li, X., et al., Effect of different Agrobacterium rhizogenes strains on hairy root induction and phenylpropanoid biosynthesis in Tartary buckwheat (Fagopyrum tataricum Gaertn), Front. Microbiol., 2016, vol. 7, p. 318. https://doi.org/10.3389/fmicb.2016.00318

    Article  PubMed  PubMed Central  Google Scholar 

  21. Tiwari, R.K., Trivedi, M., Guang, Z.C., et al., Agrobacterium rhizogenes mediated transformation of Scutellaria baicalensis and production of flavonoids in hairy roots, Biol. Plant., 2008, vol. 52, p. 26. https://doi.org/10.1007/s10535-008-0004-9

    Article  CAS  Google Scholar 

  22. Wang, X., Ding, G., Liu, B., and Wang, Q., Flavonoids and antioxidant activity of rare and endangered fern: Isoetes sinensis, PLoS One, 2020, vol. 15, no. 5, e0232185. https://doi.org/10.1371/journal.pone.0232185

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Zhang, H.C., Liu, J.M., Lu, H.Y., et al., Enhanced flavonoid production in hairy root cultures of Glycyrrhiza uralensis Fisch by combining the overexpression of chalcone isomerase gene with the elicitation treatment, Plant Cell Rep., 2009, vol. 28, pp. 1205–1213. https://doi.org/10.1007/s00299-009-0721-3

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

The study was partly supported by grant no. 2929.01/0301 of NRF of Ukraine and the grant of MES of Ukraine “Developing Methods for Deriving Active Pharmaceutical Ingredients with Anti-Inflammatory and Antioxidant Activities Based on the Biotechnology of Wormwood Roots,” 2021–2022.

Author information

Authors and Affiliations

  1. Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, 03143, Kyiv, Ukraine

    T. A. Bohdanovych, A. M. Shakhovsky, V. P. Duplij, Ya. I. Ratushnyak, M. V. Kuchuk & N. A. Matvieieva

  2. National Technical University of Ukraine–Sikorsky Kyiv Polytechnic Institute, 03056, Kyiv, Ukraine

    N. L. Poyedinok

  3. Institute of Food Biotechnology and Genomics, National Academy of Sciences of Ukraine, 04123, Kyiv, Ukraine

    N. L. Poyedinok

Authors
  1. T. A. Bohdanovych
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. M. Shakhovsky
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. V. P. Duplij
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ya. I. Ratushnyak
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. V. Kuchuk
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. N. L. Poyedinok
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. N. A. Matvieieva
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to T. A. Bohdanovych, A. M. Shakhovsky, V. P. Duplij, Ya. I. Ratushnyak, M. V. Kuchuk, N. L. Poyedinok or N. A. Matvieieva.

Ethics declarations

The authors declare that they have no conflict of interests.

This study was performed by each of the authors in compliance with ethical norms and did not include any procedures involving people or animals as objects.

Additional information

Translated by N. Tarasyuk

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bohdanovych, T.A., Shakhovsky, A.M., Duplij, V.P. et al. Effects of Genetic Transformation on the Antioxidant Activity of “Hairy” Roots of Althaea officinalis L., Artemisia vulgaris L., and Artemisia tilesii Ledeb.. Cytol. Genet. 55, 531–539 (2021). https://doi.org/10.3103/S0095452721060037

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.3103/S0095452721060037

Keywords:

Search

Navigation