| Plant latin name | Glycyrrhiza uralensis Fisher |
| Literature code | Glycyrrhiza_uralensis-Ref-1 |
| Reference | Kusano G et al., Natural Medicines 54: 199-203 (2000) |
| Summary | A strain of Glycyrrhiza uralensis, which was almost dying in the garden of Kanzo Yashiki (Enzan city, Yamanashi Prefecture) in the summer of 1992, was revived by stopping the growing with Kiwi vine (Ac tinidia chinesis ), by implantation of the stolon, and by node culture of the meristem tissue of buds |
| Objectives | Characterization and propagation of Glycyrrhiza uralensis which had beed recorded its cultivation in Enzan city, Yamanasi prefecture since 1720. |
| Materials | 1990年に甘草屋敷（山梨県塩山市上於曽）のキウイ畑に生えていたのが確認されたウラルカンゾウ．
A strain of Glycyrrhiza uralensis, which was conformed its survival in the garden of Kanzo Yashiki (Enzan city, Yamanashi Prefecture) in the summer of 1992. |
| Explant | Shoots |
| Initial culture | Shoot-tips were axenically excised under binocular microscope, inoculated on 1/3 macro salts concentration Murashige and Shoog (MS) medium supplemented with IAA 0.1 mg/l and Zeatin 0.3 mg/l（solidified with gelrite） and cultured at 25℃． |
| Shoot multiplication | Nodal segments excised from the initial shoot with 3-5 leaves were placed on 1/3 MS medium supplemented with IAA 0.1 mg/l and Zeatin 0.3 mg/l and cultured at 25℃． The plants were propagated by repeated subculture with the same cultural conditions. |
| Rooting | For rooting, the developed shoots with 5-6 leaves were cultured on 1/3 MS with IBA 0.1 mg/l． |
| Acclimation | Acclimatization in greenhouse |
| Planting | In vitro plantlet was transplated onto a pot filled with vermiculite and grewn in the greenhous. They were transplanted in to the field in autum and cultivated for 2 year. |
| Cultivation conditions | |
| Traints of regenerants | The glycyrrhizin concentration in the underground part of the field-cultivated plants for two year was 2.81%.
圃場移植後2年間栽培した再生植物体の地下部のグリチルリチン含量は，2.81%． |
| Ingredients analyzed | Glycyrrhizin |
| Extraction | After harvest of the plants, dried (at 50℃ until constant weight) and powdered glycyrrhiza was accurately weighed at 50 mg, added with 10 ml 50% ethanol containing 0.01 mg/ml p-hydroxybenzoic acid n-propyl ester as an internal standard, extracted for 20 minutes with ultrasonic. 0.45 µl of the filtered extract sulution was used as a HPLC sample. |
| Analitical methods | HPLC conditions：column Crestpak C18T(i.d. 4.6 mm x 250 mm; mobile phase acetic acid water (1→15)：acetonitrile=3:2；Flow rate 0.6 ml/min; temperature 40℃; detection UV 254 nm; injection volume 20µl． Glycyrrhizin standard was purchased by Nacalai tesque. Mean value of 3 measurements was calculated. |
| Notes | |