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Stress-Protective Effect of Furostanol Glycosides on Cultured Medicago sativa Cells in vitro under Hypothermia and Hyperosmotic Stress

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Abstract

The study of the mechanisms of adaptogenic action of furostanol glycosides (FG) on plant cells was carried out in vitro and a comparison was made of the effects of hypothermia and hyperosmotic stress on a suspension cell culture of Medicago sativa L. It has been shown that the cell population of M. sativa in vitro possesses functional specificities that determine different sensitivity to the action of these abiotic stressors, which was expressed in different levels of cell viability: high (85%) under hypothermia and low (25%) under hyperosmotic action. Hypothermia stimulated the rate of generation of superoxide anion (\({\text{O}}_{2}^{{\bullet - }}\)), and it was accompanied by a high constitutive activity of antioxidant enzymes (guaiacol-peroxidase, ascorbate peroxidase and glutathione peroxidase), the level of which reflects the compensatory potential of cells. Preservation of a high level of cell viability under the action of hypothermia, despite a 40% increase in the rate of generation of \({\text{O}}_{2}^{{\bullet - }}\), indicates that the formed reactive oxygen species did not cause damage to lipid structures and macromolecules in cells. Exogenous treatment with FG with this type of stress effect promoted an increase in the activity of antioxidant enzymes but did not have a noticeable effect on the initially high level of cell viability. In conditions of hyperosmotic stress, preliminary exposure to FG led to a threefold increase in cell survival (from 25 to 73%) and a 30% increase in the activity of soluble peroxidase in comparison with its level under the influence of only a stressor. Exposure to FG also caused an increase in activity antioxidant enzymes, a decrease in the level of lipid peroxidation and an increase in the activity of enzymes of the malate dehydrogenase (MDH) complex. However, in contrast to hypothermia, the observed changes caused a significant increase in the viability of M. sativa cells in vitro. An increase in the formation of osmolytes in the reactions NAD/NAD·H-MDH was indicated by the increased concentration of osmotic (mannitol), which causes the initial degree of cell plasmolysis, in comparison with the control. The article discusses specificities of M. sativa cell culture influencing the specificity of its stress-resistance, possible mechanisms of cell protection in vitro in hypothermia and hyperosmotic stress and the role of FG in these processes.

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REFERENCES

  1. Volkova, L.A., Maevskaya, S.N., Burgutin, A.B., and Nosov, A.M., Effect of exogenous steroid glycosides on cultured cells of potato under oxidative stress, Russ. J. Plant Physiol., 2007, vol. 54, p. 639. https://doi.org/10.1134/S1021443707050111

    Article  CAS  Google Scholar 

  2. Volkova, L.A., Urmantseva, V.V., Burgutin, A.B., Maevskaya, S.N., and Nosov, A.M., Stimulation of defense responses of in vitro potato plants by treatment with steroid glycosides under abiotic stresses, Russ. J. Plant Physiol., 2011, vol. 58, p. 928. https://doi.org/10.1134/S1021443711040236

    Article  CAS  Google Scholar 

  3. Volkova, L.A., Urmantseva, V.V., Burgutin, A.B., and Nosov, A.M., Characteristics of eliciting effects of furostanol glycosides on cultured yam cells, Russ. J. Plant Physiol., 2018, vol. 65, p. 427. https://doi.org/10.1134/S1021443718020085

    Article  CAS  Google Scholar 

  4. Volkova, L.A., Urmantseva, V.V., Burgutin, A.B., and Nosov, A.M., Sensitivity of antioxidant status of plant cells to furostanol glycosides, Russ. J. Plant Physiol., 2016, vol. 63, p. 784. https://doi.org/10.7868/S0015330316060154

    Article  Google Scholar 

  5. Volkova, L.A., Urmantseva, V.V., Burgutin, A.B., and Nosov, A.M., Effect of furostanol glycosides from Dioscorea deltoidea on redox state of alfalfa cells in vitro, Russ. J. Plant Physiol., 2021, vol. 68, p. 1098. https://doi.org/10.1134/S102144372105023X

    Article  CAS  Google Scholar 

  6. Ogasawara, Y., Kaya, H., Hiraoka, G., Yumoto, F., Kimura, S., Kadota, Y., Hishinuma, H., Senzaki, E., Yamagoe, S., Nagata, K., and Nara, M., Synergistic activation of the Arabidopsis NADPH oxidase AtrbohD by Ca2+ and phosphorylation, J. Biol. Chem., 2008, vol. 283, p. 8885. https://doi.org/10.1074/jbc.M708106200

    Article  CAS  PubMed  Google Scholar 

  7. Scandalios, J.G., Oxidative stress: molecular perception and transduction of signals triggering antioxidant gene defenses, Bras. J. Med. Biol. Res., 2005, vol. 38, p. 995. https://doi.org/10.1590/s0100-879x2005000700003

    Article  CAS  Google Scholar 

  8. Theocharis, A., Clement, C., and Barka, E.A., Physiological and molecular changes in plants grown at low temperatures, Planta, 2012, vol. 235, p. 1091. https://doi.org/10.1007/s00425-012-1641-y

    Article  CAS  PubMed  Google Scholar 

  9. Passardi, F., Cosio, C., Penel, C., and Dunand, C., Peroxidases have more functions than a Swiss army knife, Plant Cell Rep., 2005, vol. 24, p. 255. https://doi.org/10.1007/s00299-005-0972-6

    Article  CAS  PubMed  Google Scholar 

  10. Ackermann, F. and Stanislas, T., The plasma membrane—an integrating compartment for mechano-signaling, Plants, 2020, vol. 9, p. 505. https://doi.org/10.3390/plants9040505

    Article  CAS  PubMed Central  Google Scholar 

  11. Volkova, L.A., Urmantseva, V.V., Popova, E.V., and Nosov, A.M., Physiological, cytological and biochemical stability of Medicago sativa L. cell culture after 27 years of cryogenic storage, CryoLetters, 2015, vol. 36, p. 252.

    CAS  PubMed  Google Scholar 

  12. Lin, C.C. and Kao, C.H., NaCl induced changes in ionically bound peroxidase activity in roots of rice seedlings, Plant Soil, 1999, vol. 216, p. 147. https://doi.org/10.1023/A:1004714506156

    Article  CAS  Google Scholar 

  13. Beauchamp, C. and Fridovich, J., Superoxide dismutase: improved assays and an assay applicable to acrylamide gels, Anal. Biochem., 1971, vol. 44, p. 276.

    Article  CAS  Google Scholar 

  14. Shul’gin, K.K., Popov, S.S., Rakhmanova, T.I., Popova, T.N., Safonova, O.A., Verevkin, A.N., Semenikhina, A.V., and Goncharova, E.I., Glutathione peroxidase activity during liver dysfunction and chromatographic isolation of enzyme to study regulatory properties, Sorbtsionnye Khromatogr. Protsessy, 2016, vol. 16, p. 916.

    Google Scholar 

  15. Gerbling, K.-P., Kelly, G.J., Fischer, K.-H., and Latzko, E., Partial purification and properties of soluble ascorbate peroxidases from pea leaves, J. Plant Physiol., 1984, vol. 115, p. 59.

    Article  CAS  Google Scholar 

  16. Liszkay, A., van der Zalm, E., and Schopfer, P., Production of reactive oxygen intermediates (O2•–, H2O2, and OH) by maize roots and their role in wall loosening and elongation growth, Plant Physiol., 2004, vol. 136, p. 3114. https://doi.org/10.1104/pp.104.044784

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Heath, R.L. and Packer, L., Photoperoxidation in isolated chloroplasts. 1. Kinetics and stoichiometry of fatty acid peroxidation, Arch. Biochem. Biophys., 1968, vol. 125, p. 189.

    Article  CAS  Google Scholar 

  18. Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J., Protein measurement with Folin phenol reagent, J. Biol. Chem., 1951, vol. 193, p. 265.

    Article  CAS  Google Scholar 

  19. Chinnusamy, V., Zhu, J., and Zhu, J.-K., Cold stress regulation of gene expression in plants, Trends Plant Sci., 2007, vol. 12, p. 444. https://doi.org/10.1016/j.tplants.2007.07.002

    Article  CAS  PubMed  Google Scholar 

  20. Kawano, T., Roles of the reactive oxygen species generating peroxidase reactions in plant defense and growth induction, Plant Cell Rep., 2003, vol. 21, p. 829. https://doi.org/10.1007/s00299-003-0591-z

    Article  CAS  PubMed  Google Scholar 

  21. Tanaka, K., Takio, S., Yamamoto, I., and Satoh, T., Purification of the cytosolic CuZn-superoxide dismutase (CuZn-SOD) of Marchantia paleacea var. diptera and its resemblance to CuZn-SOD from chloroplasts, Plant Cell Physiol., 1996, vol. 37, p. 523.

    Article  CAS  Google Scholar 

  22. Foyer, C.H. and Noctor, G., Defining robust redox signaling within the context of the plant cell, Plant Cell Environ., 2015, vol. 38, p. 239. https://doi.org/10.1111/pce.13659

    Article  CAS  PubMed  Google Scholar 

  23. Mehla, N., Sindhi, V., Josula, D., Bisht, P., and Wani, S.H., An introduction to antioxidants and their roles in plant stress tolerance, in Reactive Oxygen Species and Antioxidant Systems in Plants: Role and Regulation under Abiotic Stress, Khan, M.I.R. and Khan, N.A., Eds., Singapore: Springer-Verlag, 2017, p. 1. https://doi.org/10.1007/978-981-10-5254-5

  24. Polesskaya, O.G., Rastitel’naya kletka i aktivnye formy kisloroda (Plant Cell and Reactive Oxygen Species), Moscow: KDU, 2007.

  25. Kolupaev, Yu.E., Plant cell antioxidants and their role in ROS signaling and plant resistance, Usp. Sovrem. Bio-l., 2016, vol. 136, p. 181.

    Google Scholar 

  26. Volkova, L.A., Urmantseva, V.V., Klyushin, A.G., Burgutin, A.B., and Nosov, A.M., Activity of respiratory pathways in cultured yam cells under the influence of furostanol glycosides, Russ. J. Plant Physiol., 2020, vol. 67, p. 344. https://doi.org/10.1134/S1021443715040172

    Article  CAS  Google Scholar 

  27. Volinsky, R. and Kinnunen, P.K.J., Oxidized phosphatidylcholines in membrane-level cellular signaling: from biophysics to physiology and molecular pathology, FEBS J., 2013, vol. 280, p. 2806. https://doi.org/10.1111/febs.12247

    Article  CAS  PubMed  Google Scholar 

  28. Evstigneev, M.P., Zav’yalova, O.S., and Savchenko, E.V., Biofizika membran: uchebnoe posobie (Membrane Biophysics: Manual), Sevastopol: Sevastopol. Gos. Univ., 2019.

  29. Lokhande, V.H. and Suprasanna, P., Prospects of halophytes in understanding and managing abiotic stress tolerance, in Environmental Adaptations and Stress Tolerance of Plants in the Era of Climate Change, Ahmad, P. and Prasad, M.N.V., Eds., New York: Springer-Verlag, 2012, p. 29. https://doi.org/10.1007/978-1-4614-0815-4_2

  30. Urmantseva, V.V., Volkova, L.A., Ul’yanova, K.A., Kulichenko, I.E., and Nosov, A.M., The use of hydrogen peroxide and dicarboxylic acids to increase the osmotolerance of cell culture of Dioscorea deltoidea W., Biotekhnologiya, 2009, no. 2, p. 14.

  31. Fruhwirth, G.O., Loidl, A., and Hermetter, A. Oxidized phospholipids: from molecular properties to disease, Biochim. Biophys. Acta, Mol. Basis Dis., 2007, vol. 1772, p. 718. https://doi.org/10.1016/j.bbadis.2007.04.009

    Article  CAS  Google Scholar 

  32. Stepovaya, E.A., Shakhristova, E.V., Ryazantseva, N.V., Nosareva, O.L., Yakushina, V.D., Nosova, A.I., Gulaya, V.S., Stepanova, E.A., Chil’chigashev, R.I., and Novitsky, V.V., The role of oxidative protein modification and the glutathione system in modulation of the redox status of breast epithelial cells, Biochemistry (Moscow) Suppl. Ser. B: Biomed. Chem., 2016, vol. 10, p. 235. https://doi.org/10.1134/S1990750816030161

    Article  Google Scholar 

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This work was supported by the Russian Science Foundation, grant no. 19-14-00387.

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Correspondence to A. M. Nosov.

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Abbreviations: APO—ascorbate peroxidase; GPO—glutathione peroxidase; PO—peroxidase; SOD—superoxide dismutase; TBK-AP—active products that react with thiobarbituric acid; FG—furostanol glycosides; FA—fatty acids.

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Volkova, L.A., Urmantseva, V.V., Burgutin, A.B. et al. Stress-Protective Effect of Furostanol Glycosides on Cultured Medicago sativa Cells in vitro under Hypothermia and Hyperosmotic Stress. Russ J Plant Physiol 69, 55 (2022). https://doi.org/10.1134/S1021443722030177

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