Embryogenesis, plant regeneration and cardiac glycoside determination in Digitalis ferruginea subsp. ferruginea L

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 (GA 3 ) 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 GA 3 . 6-Benzylaminopurine (BAP) combined with a-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.

Ploidy Status, Nuclear DNA Content and Start Codon Targeted (SCoT) Genetic Homogeneity Assessment in Digitalis purpurea L., Regenerated In Vitro

“…The results are in accordance with Hesami and Dhaneshvar [ 47 ], who found that 1.5 mg/L BAP along with 0.15 mg/L IBA showed 100% shoot regeneration frequency. Contrary to our observation, BAP with NAA was proved to be an efficient combination for indirect shoot regeneration in Digitalis ferruginea [ 37 ], Rauvolfia serpentina [ 48 ] and Aspilia africana [ 49 ].…”

“…Seed dormancy is a common problem encountered in various plant species that leads to delayed fruit set formation and reduced production of viable plants. GA 3 is widely known to break seed dormancy, and it has also been used in promoting the seed germination rate under in vitro conditions in other species of Digitalis , such as D. ferruginea [ 37 ] and D. cariensis [ 8 ].…”

Influence of nutrient media on callus induction, somatic embryogenesis and plant regeneration in selected Turkish crocus species

“…In the present study, shoot regeneration was also achieved from embryogenic calli that was already cultured for somatic embryos (Data not shown; Table 2 ). However, shoot development on the callus was mediated via organogenic-like shoot meristems previously reported by Verma et al [51] . Therefore, it can not be ruled out that, next to somatic embryogenesis, adventitious shoot formation was also involved in the plant propagation [51] .…”

The Foxgloves (Digitalis) Revisited

“…Digoxin and digitoxin were not detected in either source [69]. Efficient in vitro plant regeneration protocols for Digitalis ferruginea were also reported [70].…”