Temperature stress response of althaea officinalis «hairy» root lines carrying human interferon α2b gene

Matvieieva N.A., Ratushnyak Y.I., Duplij V.P., Shakhovsky A.M., Kuchuk M.V.

SUMMARY. «Hairy» roots, obtained by genetic transformation of plants using soil phytopathogenic bacteria Agrobacterium rhizogenes, are valuable producers of important se-condary metabolites with medicinal properties and a
convenient model object for studying the response of plants to adverse environmental conditions. This paper compares the postponed response of «hairy» roots of Althaea officinalis L. to the short-term cold and high-temperature stresses. The results indicate that the «hairy» roots of different lines of A. officinalis (individual transformational events) differ in sensitivity to short-term temperature stress, regardless of the vector used for the transformation and the presence of human interferon ifn-α2b gene. The high temperature led to significant inhibition of root growth of all lines, except the one that had the highest content of flavonoids under the control conditions. On the other hand, short-term cultivation of «hairy» roots at low temperature did not cause growth inhibition. Simultaneously with the inhibition of growth by high temperature conditions, an increase in the synthesis of flavonoids was observed. Probably, it was a response of the roots to the action of high temperature as a stress factor. The strong (R² = 0,78) linear dependence between antioxidant activity of «hairy» root extracts and the total flavonoid content was determined. Thus, flavonoids synthesized in A. officinalis «hairy» roots may be involved in the process of response and adaptation of the roots to high temperature stress.

Key words: Agrobacterium rhizogenes, Althaea officinalis, “hair” roots, temperature stress, flavonoids, antioxidant activity

Tsitologiya i Genetika 2021, vol. 55, no. 3, pp. 3-9

• Institute of Cell Biology and Genetic Engineering of National Academy of Sciences of Ukraine, 148 Academika Zabolotnoho St., 03143, Kyiv, Ukraine

E-mail: duplijv icbge.org.ua

Matvieieva N.A., Ratushnyak Y.I., Duplij V.P., Shakhovsky A.M., Kuchuk M.V. Temperature stress response of althaea officinalis «hairy» root lines carrying human interferon α2b gene, Tsitol Genet., 2021, vol. 55, no. 3, pp. 3-9.

In "Cytology and Genetics":
N. A. Matvieieva, Y. I. Ratushnyak, V. P. Duplij, A. M. Shakhovsky & M. V. Kuchuk Effect of Temperature Stress on the Althaea officinalis’s “Hairy” Roots Carrying the Human Interferon α2b Gene, Cytol Genet., 2021, vol. 55, no. 3, pp. 207–212
DOI: 10.3103/S0095452721030051

References

1. Agati, G., Azzarello, E., Pollastri, S., and Tattini, M., Flavonoids as antioxidants in plants: location and functional significance, Plant Sci., 2012, vol. 196, pp. 67–76.

2. Boo, H.O., Chon, S.U., and Lee, S.Y., Effects of temperature and plant growth regulators on anthocyanin synthesis and phenylalanine ammonia-lyase activity in chicory (Cichorium intybus L.), J. Hortic. Sci. Biotechnol., 2006, vol. 81, pp. 478–482. https://doi.org/10.1080/14620316.200.11512091

3. Choi, S., Kwon, Y.R., Hossain, M.A., et al., A mutation in ELA1, an age-dependent negative regulator of PAP1/MYB75, causes UV- and cold stress-tolerance in Arabidopsis thaliana seedlings, Plant Sci., 2009, vol. 176, pp. 678– 686. https://doi.org/10.1016/j.plantsci.2009.02.010

4. Fini, A., Brunetti, C., Di Ferdinando, M., et al., Stress-induced flavonoid biosynthesis and the antioxidant machinery of plants, Plant Signal. Behav., 2011, vol. 6, pp. 709–711. https://doi.org/10.4161/psb.6.5.15069

5. Havryliuk, O., Matvieieva, N., Tashyrev, O., and Yastremskaya, L., Influence of cold stress on growth and flavonoids accumulation in Artemisia tilesii “hairy” root culture, in Agrobiodiversity for Improving Nutrition, Health and Life Quality, 2017, pp 163–167.

6. 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

7. Matvieieva, N.A., Generation of Tragopogon porrifolius and Althaea officinalis “hairy” roots using Agrobacterium rhizogenes, Bull. Vavilov Soc. Genet. Breeders Ukr., 2012, vol. 10, pp. 262–268.

8. Matvieieva, N.A., Kishchenko, O.M., Potrochov, A.O., et al., Regeneration of transgenic plants from hairy roots of Cichorium intybus L. var. Foliosum Hegi, Cytol. Genet., 2011, vol. 45, pp. 277–281. https://doi.org/10.3103/S0095452711050082

9. Matvieieva, N.A., Morgun, B.V., Lakhneko, O.R., 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

10. Matvieieva, N.A., Shachovsky, A.M., Gerasymenko, I.M., et al., Agrobacterium-mediated transformation of Cichorium intybus L. with interferon-α2b gene, Biopolym. Cell, 2009, vol. 25, pp. 120–125. https://doi.org/10.7124/bc.0007D4

11. Murashige, T. and Skoog, F., A revised medium for rapid growth and bio assays with tobacco tissue cultures, Physiol. Plant., 1962, vol. 15, pp. 473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

12. 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

13. Ramakrishna, A. and Ravishankar, G.A., Influence of abiotic stress signals on secondary metabolites in plants, Plant Signal. Behav., 2011, vol. 6, pp. 1720–1731.

14. Sanghera, G.S., Wani, S.H., Hussain, W., and Singh, N.B., Engineering cold stress tolerance in crop plants, Curr. Genomics, 2011, vol. 12, pp. 30–43. https://doi.org/10.2174/138920211794520178

15. Schulz, E., Tohge, T., Zuther, E., et al., Flavonoids are determinants of freezing tolerance and cold acclimation in Arabidopsis thaliana, Sci. Rep., 2016, vol. 6, art. 34027. https://doi.org/10.1038/srep34027

16. Schulz, E., Tohge, T., Zuther, E., et al., Natural variation in flavonol and anthocyanin metabolism during cold acclimation in Arabidopsis thaliana accessions, Plant Cell Environ., 2015, vol. 38, pp. 1658–1672. https://doi.org/10.1111/pce.12518

17. Shamloo, M., Babawale, E.A., Furtado, A., et al., Effects of genotype and temperature on accumulation of plant secondary metabolites in Canadian and Australian wheat grown under controlled environments, Sci. Rep., 2017, vol. 7, art. 9133. https://doi.org/10.1038/s41598-017-09681-5

18. Srivastava, S. and Srivastava, A.K., Hairy root culture for mass-production of high-value secondary metabolites, Crit. Rev. Biotechnol., 2007, vol. 27, pp. 29–43. https://doi.org/10.1080/07388550601173918

19. Wahid, A., Physiological implications of metabolite biosynthesis for net assimilation and heat-stress tolerance of sugarcane (Saccharum officinarum) sprouts, J. Plant Res., 2007, vol. 120, pp. 219–228. https://doi.org/10.1007/s10265-006-0040-5

20. Wang, L., Tu, Y.-C., Lian, T.-W., et al., Distinctive antioxidant and antiinflammatory effects of flavonols, J. Agric. Food Chem., 2006, vol. 54, pp. 9798–9804.https://doi.org/10.1021/jf0620719

21. Wang, S.Y. and Zheng, W., Effect of plant growth temperature on antioxidant capacity in strawberry, J. Agric. Food Chem., 2001, vol. 49, pp. 4977–4982. https://doi.org/10.1021/jf0106244

22. Wu, G., Johnson, S.K., Bornman, J.F., et al., Growth temperature and genotype both play important roles in sorghum grain phenolic composition, Sci. Rep., 2016, vol. 6, art. 21835. https://doi.org/10.1038/srep21835