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Effect of Auxins and Cytokinins on Growth and Biosynthetic Characteristics of Suspension Cell Culture of Tribulus terrestris L.

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Abstract

The work aimed on studying the effect of synthetic phytohormones on growth and biosynthetic parameters of a suspension culture of Tribulus terrestris L., and the influence of cytokinins on these characteristics. In addition to the previously obtained results about alternative action of two types of auxins, 2,4-D and α-NAA (the first of them activates cell growth in vitro but inhibits the formation of steroidal glycosides, while the second causes cytodifferentiation, leading to a slowdown and arrest of culture growth, but activates the formation of secondary compounds), it was shown that the type of cytokinin and its concentration can modulate its action. It was found that, after transferring a culture from a “growth” medium (containing 2,4-D) to a medium with α-NAA and a different composition of cytokinins (BAP or kinetin in various concentrations), it retains the ability to actively grow for at least three growing cycles. Then, with a suboptimal combination of phytohormones, the cell culture stops growing and dies. However, with a certain composition of growth regulators in a nutrient medium (1 or 2 mg/L α-NAA + 1 mg/L kinetin), after a long lag phase (12–18 days), culture growth resumes and its long-term cultivation is subsequently possible. Suspension on a medium with 2 mg/L α-NAA and 1 mg/L kinetin T. terrestris were maintained for 70 growing cycles without deterioration of growth characteristics (the maximum level of accumulation of dry biomass Mmax was 12–16 g/L, growth index I was 13–18, specific growth rate μ was 0.18–0.28 days–1). In this case, the content of steroidal glycosides in cells did not fall below 0.2–0.3% of dry biomass. Using the UPLC-ESI-MS method, the presence in the cell biomass of at least four steroidal glycosides of the furostanol type—derivatives of hydroxy-diosgenin and tigogenin/neotigogenin—was shown. The results obtained confirm the general patterns of development of cell populations in vitro and are important for optimizing the growth and biosynthetic characteristics of plant cell cultures.

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Funding

The work on the cultivation of cell cultures was carried out with the financial support of a grant from the Russian Science Foundation, project no. 19-14-00387; phytochemical analyzes were carried out on the basis of the “Scientific and Production Biotechnological Complex for the Study, Preservation, and Practical Application of Cultured Cells and Organs of Higher Plants and Microalgae” with financial support from the Megagrant of the Government of the Russian Federation (agreement no. 075-15-2019-1882).

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Authors and Affiliations

  1. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, Moscow, Russia

    A. G. Klyushin, S. V. Tomilova, D. V. Kochkin & A. M. Nosov

  2. Moscow State University, Moscow, Russia

    D. V. Kochkin & A. M. Nosov

  3. Ural Federal University, Yekaterinburg, Russia

    B. A. Galishev

Authors
  1. A. G. Klyushin
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  2. S. V. Tomilova
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  3. D. V. Kochkin
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  4. B. A. Galishev
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  5. A. M. Nosov
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Correspondence to A. G. Klyushin or A. M. Nosov.

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Conflict of interests. The authors declare that they have no conflicts of interest.

Statement on the welfare of humans or animals. This article does not contain any studies involving humans or animals performed by any of the authors.

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Abbreviations: FG—furostanol glycosides; UPLC-ESI-MS—ultrahigh-performance liquid chromatography coupled with electrospray ionization mass spectrometry.

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Klyushin, A.G., Tomilova, S.V., Kochkin, D.V. et al. Effect of Auxins and Cytokinins on Growth and Biosynthetic Characteristics of Suspension Cell Culture of Tribulus terrestris L.. Russ J Plant Physiol 69, 52 (2022). https://doi.org/10.1134/S1021443722020078

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  • DOI: https://doi.org/10.1134/S1021443722020078

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