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Obtaining and Study of Callus and Suspension Plant Cell Cultures of Tribulus terrestris L., a Producer of Steroidal Glycosides

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Abstract

Callus and suspension plant cell cultures of Tribulus terrestris L., a valuable medicinal plant producing steroidal glycosides, were obtained. The seeds from an American population of T. terrestris were used as explants. Regulation of the production and growth of cell cultures, as well as the biosynthetic characteristics of the cell lines, were studied. The combination of phytohormones of 2,4-D (2.0 mg/L) and BAP (1.0 mg/L) was found to be optimal for callus induction and cultivation. Suspension cell culture obtained in liquid medium of the same composition showed such high growth characteristics during prolonged cultivation (more than 2 years) as a maximum accumulation of dry biomass of 13 g/L, specific growth rate at exponential phase of 0.24 day–1, and economical coefficient of 0.39. A semicontinuous mode of cultivation was used to grow the plant cell suspension in a lab-scale bioreactor. Screening of the steroidal glycosides in the obtained cell cultures was carried out. Steroidal glycosides were not found in the callus cultures. However, as was demonstrated by TLC and UPLC ESI MS methods, the suspension culture contained furostanol glycosides, and their amount increased during the cultivation process. These results support the hypothesis of the autoselection of cultivated cells containing compounds promoting their proliferation in vitro.

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Abbreviations

BAP:

6-benzylaminopurine

2,4-D-2:

2,4-dichlorophenoxyacetic acid

NAA:

α-naphthylacetic acid

TLC:

thin layer chromatography

UPLC ESI MS:

ultra performance liquid chromatography-electrospray ionization-tandem mass spectrometry

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Authors and Affiliations

  1. Lomonosov Moscow State University, Moscow, 119234, Russia

    M. T. Khandy, D. V. Kochkin, E. S. Sukhanova & A. M. Nosov

  2. Ammosov Northeastern Federal University, Yakutsk, 677000, Russia

    M. T. Khandy

  3. Timiryazev Institute of Plant Physiology, Russian Academy of Science, Moscow, 127276, Russia

    D. V. Kochkin, E. S. Sukhanova, A. G. Klyushin, I. M. Ivanov & A. M. Nosov

  4. Yeltsin Ural Federal University, Yekaterinburg, 620002, Russia

    S. V. Tomilova & B. A. Galishev

Authors
  1. M. T. Khandy
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  2. D. V. Kochkin
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  3. S. V. Tomilova
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  4. B. A. Galishev
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  5. E. S. Sukhanova
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  6. A. G. Klyushin
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  7. I. M. Ivanov
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  8. A. M. Nosov
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Correspondence to M. T. Khandy.

Additional information

Original Russian Text © M.T. Khandy, D.V. Kochkin, S.V. Tomilova, B.A. Galishev, E.S. Sukhanova, A.G. Klyushin, I.M. Ivanov, A.M. Nosov, 2016, published in Biotekhnologiya, 2016, Vol. 32, No. 4, pp. 21–30.

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Khandy, M.T., Kochkin, D.V., Tomilova, S.V. et al. Obtaining and Study of Callus and Suspension Plant Cell Cultures of Tribulus terrestris L., a Producer of Steroidal Glycosides. Appl Biochem Microbiol 53, 800–806 (2017). https://doi.org/10.1134/S0003683817080038

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  • DOI: https://doi.org/10.1134/S0003683817080038

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