Abstract
The present study reports, for the first time, an efficient in vitro plant regeneration protocol for Digitalis ferruginea subsp. ferruginea L. (rusty foxglove). We have used different concentrations of gibberellic acid (GA3) on Murashige and Skoog (MS) medium to assess the germination frequency of seeds. High frequency of germination was achieved on MS medium with 1.0 mg l−1 GA3. 6-Benzylaminopurine (BAP) combined with α-naphtaleneacetic acid (NAA) or 2, 4-dichlorophenoxy acetic acid (2, 4-D) in the induction MS medium induced both somatic embryogensis and shoot organogenesis. The highest percentage of callus growth (85 %) was obtained when hypocotyl explants were cultured on MS medium containing 0.5 mg l−1 2, 4-D plus 1.0 mg l−1 BAP. The maximum mean number of somatic embryos (7.3 ± 1.3 embryos) or shoots (12.0 ± 1.1 shoots) per callus was obtained when medium contained 0.25 mg l−1 NAA plus 1.0 mg l−1 BAP or 0.5 mg l−1 NAA plus 2.0 mg l−1 BAP. The regenerated shoots easily rooted on MS medium. Higher amounts of lanatoside C [13.2 ± 0.5 mg 100 g−1 dry weight (dw)] and digoxin (2.93 ± 0.31 mg 100 g−1 dw) accumulation were obtained when shoots were obtained by indirect regeneration. We also investigated derivatives of cardenolides, i.e., digitoxigenin (730 ± 180 mg 100 g−1 dw), gitoxigenin (50 ± 20 mg 100 g−1 dw) and digoxigenin (490 ± 170 mg 100 g−1 dw) from natural samples.
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Abbreviations
- BAP:
-
6-Benzylaminopurine
- GA3 :
-
Gibberellic acid
- HPLC:
-
High performance liquid chromatography
- IAA:
-
Indole-3-acetic acid
- IBA:
-
Indole-3-butyric acid
- NAA:
-
α-Naphthalene acetic acid
- PGR:
-
Plant growth regulator
- TDZ:
-
Thidiazuron
- 2, 4-D:
-
2, 4-Dichlorophenoxy acetic acid
- MS:
-
Murashige and Skoog
- MSO:
-
MS medium without hormones
- TLC:
-
Thin layer chromatography
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Acknowledgments
The authors thanks to TÜBİTAK for financial support (project no: TOVAG-106O470), a research grant for the international research fellowship programme (2214) to B.Y. and the Turkish Government for providing scholarship to S.K.Verma through the MHRD, India. Special thanks to Prof. Dr. W. Kreis (Friedrich-Alexander-Universität Erlangen-Nürnberg, Germany) for their technical support towards cardenolide analyses (genine group). The authors express their sincere thanks to Prof. Sergio J. Ochatt for his critical reading of this manuscript.
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Verma, S.K., Yucesan, B., Sahin, G. et al. Embryogenesis, plant regeneration and cardiac glycoside determination in Digitalis ferruginea subsp. ferruginea L. Plant Cell Tiss Organ Cult 119, 625–634 (2014). https://doi.org/10.1007/s11240-014-0562-9
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DOI: https://doi.org/10.1007/s11240-014-0562-9