ISSN 0564-3783  



Main page
Contacts
Themes
Archive  
Themes
Subscription
Information to authors
Editorial board
Mobile version


In Ukrainian

Export citations
UNIMARC
BibTeX
RIS





Effect of the genetic transformation on the antioxidant activity of Althaea officinalis L., Artemisia vulgaris L. and Artemisia tilesii Ledeb. hairy roots

Bohdanovyh T., Shakhovsky A., Duplij V., Ratushnyak Y., Kuchuk M., Poyedinok N., Matvieieva N.

 




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 58 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

  1. Institute of Cell Biology and Genetic Engineering of National Academy of Sciences of Ukraine, 148 Academika Zabolotnoho St., 03143, Kyiv, Ukraine
  2. National Technical University of Ukraine Igor Sikorsky Kyiv Polytechnic Institute, 37, Prosp. Peremohy, Kyiv, 03056, Ukraine
  3. Institute of Food Biotechnology and Genomics of National Academy of Sciences of Ukraine, 2a Osypovskoho St., 04123, Kyiv, Ukraine

E-mail: bogdanovych_tais ukr.net, an48sha gmail.com, duplijv icbge.org.ua, yakivr yahoo.com, nkuchuk icbge.org.ua, poyedinok ukr.net, joyna56 gmail.com

Bohdanovyh T., Shakhovsky A., Duplij V., Ratushnyak Y., Kuchuk M., Poyedinok N., Matvieieva N. Effect of the genetic transformation on the antioxidant activity of Althaea officinalis L., Artemisia vulgaris L. and Artemisia tilesii Ledeb. hairy roots, Tsitol Genet., 2021, vol. 55, no. 6, pp. 40-50.

In "Cytology and Genetics":
T. A. Bohdanovych, A. M. Shakhovsky, V. P. Duplij, Ya. I. Ratushnyak, M. V. Kuchuk, N. L. Poyedinok & N. A. Matvieieva Effects of Genetic Transformation on the Antioxidant Activity of Hairy Roots of Althaea officinalis L., Artemisia vulgaris L., and Artemisia tilesii Ledeb., Cytol Genet., 2021, vol. 55, no. 6, pp. 531539
DOI: 10.3103/S0095452721060037


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. 13711381. 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

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. 787797. https://doi.org/10.1007/s12298-019-00669-1

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. 3544. https://doi.org/10.1016/j.plaphy.2020.01.002

5. Giri, A. and Narasu, M.L., Transgenic hairy roots: recent trends and applications, Biotechnol. Adv., 2000, vol. 18, pp. 122. https://doi.org/10.1016/S0734-9750(99)00016-6

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. 437448. https://doi.org/10.1556/018.69.2018.4.6

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

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. 281288. https://doi.org/10.1007/s12298-019-00753-6

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

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. 342345 https://doi.org/10.1080/10826068.2015.1031393

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. 8287. https://doi.org/10.1080/10826068.2018.1536994

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. 177183. https://doi.org/10.1016/j.plaphy.2020.04.020

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. 619633.https://doi.org/10.1007/s11240-019-01757-6

14. Pekal, A. and Pyrzynska, K., Evaluation of aluminium complexation reaction for flavonoid content assay, Food Anal. Methods, 2014, vol. 7, pp. 17761782. https://doi.org/10.1007/s12161-014-9814-x

15. Pietta, P.G., Flavonoids as antioxidants, J. Nat. Prod., 2000, vol. 63, pp. 10351042. https://doi.org/10.1021/np9904509

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. 427442. 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. 29722979. https://doi.org/10.1016/j.sjbs.2020.08.050

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. 25882594. https://doi.org/10.1021/jf405695y

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

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

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

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. 12051213. https://doi.org/10.1007/s00299-009-0721-3

Copyright© ICBGE 2002-2022 Coded & Designed by Volodymyr Duplij Modified 17.08.22