Abstract
Callus and suspension cell cultures of rare foxglove species, Digitalis ciliata and D. grandiflora, were induced from cotyledons and hypocotyls of in vitro seedlings, and their growth and phytochemical profiles were investigated. In both species, callus induction was more efficient from leaf explants (60–80%) than from hypocotyl explants (15–35%). Callus cultures of both species grew well with growth indices ranged from 5 to 10 depending on the genotype. Suspension culture growth profiles also differed between the two species with a 10–11-days lag-phase observed for D. grandiflora and a bi-phasic growth curve without lag-phase recorded for D. ciliata. The main growth characteristics of the D. grandiflora suspension cell culture (maximum biomass accumulation ~ 14 g/L, growth index ~ 10, economic coefficient ~ 0.42, biomass productivity ~ 0.53 g/(L × day)) were 1.5–3 times higher than those for D. ciliata. Ten compounds were identified in cell biomass using UPLC-ESI-Q-TOF-MS: phenylethanoids digiciliside A, digiciliside B, maxoside, purpureaside E, their methyl derivatives and isomers, and two furostanol glycosides with aglycone tigogenin. Phenylethanoid glycosides were major compounds and comprised 0.8–1.1% of dry weight. During the two-years cultivation, suspension cultures retained the ability to accumulate most of the identified compounds evidencing for stability of species-specific secondary metabolism in cultured foxglove cells during this period.
Key message
Phenylethanoid glycosides (1.1% DW) and steroidal furostanol glycosides were identified in callus and suspension cell cultures of rare medicinal species Digitalis ciliata and D. grandiflora developed for the first time.
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Abbreviations
- α-NAA:
-
α-Naphthylacetic acid
- BAP:
-
6-Benzylaminopurine
- HPLC-ESI-MS:
-
High-performance liquid chromatography coupled with electrospray ionization mass spectrometry
- MS:
-
Murashige and Skoog medium
- SIM:
-
Selected ion monitoring
- UPLC-ESI-Q-TOF-MS:
-
Ultrahigh-performance liquid chromatography coupled with electrospray ionization hybrid quadrupole time-of-flight mass spectrometry
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Acknowledgements
The research on development and maintenance of cell cultures was financially supported by the Ministry of Science and Higher Education of Russian Federation through Megagrant project no. 075-15-2019-1882 and performed using the equipment of the large-scale research facilities “All-Russian Collection of cell cultures of higher plants” and “Experimental biotechnological facility” of the Institute of Plant Physiology of Russian Academy of Sciences. The results of cell culture growth assessment and phytochemical analysis were obtained within the state assignment of Ministry of Science and Higher Education of the Russian Federation (themes no. 121041200194-7 and 121050500047-5, respectively).
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SVT—development and cultivation of callus and suspension cell cultures, determination of growth characteristics, preparation of extracts for quantitative phytochemical analysis, manuscript writing and editing; DVK—qualitative and quantitative phytochemical analysis, manuscript writing and editing; TMT—preparation of extracts for qualitative phytochemical analysis; ESG—development and cultivation of callus cell cultures, manuscript editing; EAL—development of callus cell cultures; BAG—qualitative phytochemical analysis; AMN - experiment planning and supervision, fund acquisition, manuscript writing and editing.
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Tomilova, S.V., Kochkin, D.V., Tyurina, T.M. et al. Growth and biosynthetic profiles of callus and suspension cell cultures of two rare foxglove species, Digitalis grandiflora Mill. and D. ciliata Trautv.. Plant Cell Tiss Organ Cult 149, 213–224 (2022). https://doi.org/10.1007/s11240-022-02271-y
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DOI: https://doi.org/10.1007/s11240-022-02271-y