| Plant latin name | Glycyrrhiza uralensis Fisher |
| Literature code | Glycyrrhiza_uralensis-Ref-2 |
| Reference | Wongwicha W et al., Z Naturforsch, C, 63: 413–417 (2008) |
| Summary | Licorice plants, Glycyrrhiza glabra, G. uralensis, and G. inflata, were investigated for callus induction using Murashige and Skoog (MS) medium combined with auxins and cytokinins. After 4 weeks of culture, 33-100% of leaf or stem explants formed calli. Maximum of shoot induction from callus cultures was achieved by G. inflata stem explants cultured on MS medium supplemented with 1 mg/l α-naphthaleneacetic acid (NAA) and 0.5 mg/l 6-benzyladenine (BA) (67%) which also gave maximum shoot formation per explant (two shoots per explant). These results indicated that all three Glycyrrhiza species regenerated shoots from callus cultures on MS medium combined with NAA and BA or only thidiazuron (TDZ; 0.1 and 0.5 mg/l). Glycyrrhizin contents of G. uralensis calli induced using MS medium in combination with NAA and BA [(27.60±8.47) μg/g DW] or TDZ alone [(36.52 ± 2.45) μg/g DW] were higher than those found in other combinations. |
| Objectives | Investigation on the efficiency of callus induction and development of a regeneration system in three licorice plants, Glycyrrhiza glabra, G. uralensis and G. inflata |
| Materials | G. glabra, G. uralensis and G. inflata seeds obtained from the Institute of Botany, Mongolian
Academy of Sciences, Ulaanbaatar, Mongolia. |
| Explant | G. glabra, G. uralensis and G. inflata seeds obtained from the Institute of Botany, Mongolian
Academy of Sciences, Ulaanbaatar, Mongolia. |
| Initial culture | The seeds were washed with sterile distilled water and were surface-sterilized in 10% sodium hypochlorite for 15-20 min. After being washed three times with sterilized water, the seeds were immersed in 70% ethanol for 1 min and then germinated on hormone-free Murashige and Skoog (MS) medium containing 3% sucrose (w/v), pH 5.5. Germination started within 5 d and was carried out at (25±1)℃ under 16 h light/day. Plantlets were subcultured on the same medium every 4 weeks. |
| Shoot multiplication | To evaluate callus induction, leaf and stem segments (0.5 cm) from 2 weeks fully grown in vitro plantlets were cultured on MS medium (3% sucrose, 0.9% agar) with growth regulators, i. e. combinations of NAA (0.5-1 mg/l), 2,4-D (0.5-1 mg/l), BA(0.5-1 mg/l) and Kin (0.5-1mg/l), and TDZ alone (0.1-1 mg/l) at a light intensity of 70W/m2, 16h/day. After 4 weeks, the initial calli were induced (33-100%) and subcultured on the same medium. Shoot formation from a callus was observed with showing maximum of shoot induction from callus cultures achieved by G. inflata stem explants cultured on MS medium supplemented with 1 mg/l α-naphthaleneacetic acid (NAA) and 0.5 mg/l 6-benzyladenine (BA) (67%) which also gave maximum shoot formation per explant (two shoots per explant). All three Glycyrrhiza species regenerated shoots from callus cultures on MS medium combined with NAA and BA or only thidiazuron (TDZ; 0.1 and 0.5 mg/l)(best shoot induction % G. uralensis: TDZ0.1 58.33%, G. glabra: TDZ0.1 or NAA0.5+BA1 50.0%, G. inflata: NAA1+BA0.5 66.67%). This is the first report regarding the shoot formation from a callus culture of licorice plants. |
| Rooting | The initial regenerated shoots were subcultured after 4 weeks
of callus induction on MS medium without hormones. After being subcultured for 4 weeks, the regenerated shoots were rooted. |
| Acclimation | |
| Planting | |
| Cultivation conditions | |
| Traints of regenerants | |
| Ingredients analyzed | Glycyrhizin |
| Extraction | Dried samples (50 mg) of 4-week-old calli from
G. glabra, G. uralensis, and G. inflata were powdered
and extracted five times with 0.5 ml methanol
with sonication. The extracts were combined,
evaporated and then redissolved in 1 ml methanol.
The extracted solutions were diluted with 20%
methanol and glycyrrhizin analysis was performed
by competitive ELISA using anti-glycyrrhizin
MAb as described previously (Shan et al., 2001). |
| Analitical methods | |
| Notes | Shan S et al, Anal. Chem. 73: 5784-5790. |