Abstract
Hairy root cultures of Glycyrrhiza glabra producing considerable amounts of phenolic compounds were successfully established by using Agrobacterium strain C58C18(pRT GUS 104). The effect of phosphate, ammonia, nitrate and ferric-EDTA concentrations of culture medium on growth and total phenolics production of the cultures were studied. By employing statistical experimental design and linear regression analysis an improved B5 medium (B50-M) could be developed. When cultivating G. glabra hairy roots in B50-M medium we were able to obtain 9 g dried roots/l in 25 days which was twice as much as when using the initial B50 medium. According to tentative analyses the cultures did not contain glycyrrhyzin, but they produced liquiritigenin and isoliquiritigenin. The production of total phenolic substances (mg g-1 dw) was higher in the improved medium resulting in significantly higher volumetric productivity (mg phenolic compounds l-1). This will further enable the extraction and identification of the phenolic compounds produced by the cultures.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arias-Gastro C, Scragg AH, Stafford A & Rodriguez-Mendiola M (1993a) Growth charasteristics of Glycyrrhiza glabra cell suspension cultures. Plant Cell Tiss. Org. Cult. 34: 77–82
Arias-Castro C., Scragg AH & Rodriguez-Mendiola MA (1993b) The effect of cultural conditions on the accumulation of formononetin by suspension cultures of Glycyrrhiza glabra. Plant Cell Tiss. Org. Cult. 34: 63–70
Ayabe S-I, Kobayashi M, Hikichi M, Matsumoto K & Furuya T (1980) Flavonoids from the cultured cells of Glycyrrhiza echinata. Phytochemistry 19: 2179–2183
Cochran WG & Cox GM (1966) Experimental Designs. Wiley Publications, London
Demizu S, Kajiyama K, Takahashi K, Hiraga Y, Yamamoto S, Tamura Y, Okada K & Kinoshita T (1988) Antioxidant and antimicrobial constituents of licorice: Isolation and structure elucidation of a new benzofuran derivative. Chem. Pharm. Bull. 36: 3474–3479
European Pharmacopoeia (1984) Liquiritiae Radix, Licorice Root. Published under direction of Council Of Europe (pp 277/1–277/4). Maisonneuve S.A., Sainte Ruffine, France
Fenwick GR, Lutomski J & Nieman C (1990) Liquorice, Glycyrrhiza glabra L. Composition, uses and analysis. Food Chem. 38: 119–143
Flores HE, Hoy M & Pickard J (1987) Secondary metabolites from root cultures. Trends Biotechnol. 5: 64–68
Flores HE, Picard JJ & Hoy MW (1988) Production of Polyacetylenes and Thiophenes in Heterotrophic and Photosynthetic Root Cultures of Asteraceae. In: Lam SJ (Ed) Bioactive Molecules, Chemistry and Biology of Naturally occurring Acetylenes, Vol 7 (pp 233–254). Academic Press, New York
Gamborg OL (1991) Media Preparation. In: Lindsey K (Ed) Plant Tissue Culture Manual (pp 1–24). Kluwer Academic Publishers, Dordrecht
Gamborg OL, Miller RA & Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50: 151–158
Han K-H, Fleming P, Walker K, Loper M, Chilton WS, Mocek U, Gordon MP & Floss HG (1994) Genetic transformation of mature Taxus: An approach to genetically control the in vitro production of the anticancer drug, taxol. Plant Sci. 95: 187–196
Hatano T, Kavaga H, Yasuhara T & Okuda T (1988a) Two new flavonoids and other constituents in licorice root: Their relative astringency and radical scavenging effects. Chem Pharm. Bull. 36: 2090–2097
Hatano T, Yasuhara T, Miyamoto K & Okuda T (1988b) Anti-Human Immunodeficiency Virus Phenolics from Licorice. Chem. Pharm. Bull. 36: 2286–2288
Hayashi H, Fukui H & Tabata M (1988) Examination of triterpenoids produced by callus and suspension cultures of Glycyrrhiza glabra. Plant Cell Rep. 7: 508–511
Hayashi H, Fukui H & Tabata M (1993) Distribution pattern of saponins in different organs of Glycyrrhiza glabra. Planta Med. 59: 351–353.
Henry M, Edy AM, Desmarest P & Du Manoir J (1991) Glycyrrhiza glabra L. (Licorice) Cell Culture; Regeneration, and the Production of Glycyrrhizin. In: Bajaj YPS (Ed) Biotechnology in Agriculture and Forestry, Medicinal and Aromatic plants III, Vol 15 (pp 270–282). Springer-Verlag, Berlin, Heidelberg
Ko KS, Noguchi H, Ebizuka Y & Sankawa U (1989) Oligoside production by hairy root cultures transformed by Ri plasmids. Chem. Pharm. Bull. 37: 245–248
Kobayashi M, Noguchi H & Sankawa U (1985) Formation of chalcones and isoflavones by callus culture of Glycyrrhiza uralensis with different production patterns. Chem. Pharm. Bull. 33: 3811–3816
Kurz WGW & Constabel F (1985) Aspects affecting biosynthesis and biotransformation of secondary metabolites in plant cell cultures. CRC Critical Rev. Biotechnol. 2: 105–118.
Nitsch JP & Nitsch C (1969) Haploid plants from pollen grains. Science 163: 85–87
Okada K, Tamura Y, Yamamoto M, Inoue Y, Takagaki R, Takahashi K, Demizu S, Kajiyama K, Hiraga Y & Kinoshita T (1989) Identification of antimicrobial and antioxidant constituents from licorice of Russian and Xinjiang origin. Chem. Pharm. Bull. 37: 2528–2530
Parr AJ (1989) The production of secondary metabolites by plant cell cultures. J. Biotechnol. 10: 1–25
Rhodes MJC, Robins RJ, Hamill J.D, Parr A.J & Walton NJ (1987) Secondary product formation using Agrobacterium rhizogenes-transformed ‘Hairy Root’ Cultures. TCA Newsl. 53: 2–15
Saito K, Kaneko H, Yamazaki M, Yoshida M & Murakoshi I (1990) Stable transfer and expression of chimeric genes in Licorice (Glycyrrhiza uralensis) using an Ri plasmid binary vector. Plant Cell Rep. 8: 718–721
Singleton VL & Rossi JA (1965) Colorimetry of total phenolics with phosphotungstic acid reagents. Am. J. Etnol. Viticult. 16: 144–158
Toivonen L (1993) Utilization of hairy root cultures for production of secondary metabolites. Biotechnol. Progr. 9: 12–20
Vanden Berghe DA & Vlietinck AJ (1991) Screening methods for antibacterial and antiviral agents from higher plants. In: Harborne JB (Ed) Methods in Plant Biochemistry, Vol 6 (pp 47–69). Academic Press, New York
Zieslin N & Ben-Zaken R (1993) Peroxidase activity and presence of phenolic substances in peduncles of rose flowers. Plant Physiol. Biochem. 31: 333–339
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Toivonen, L., Rosenqvist, H. Establishment and growth characteristics of Glycyrrhiza glabra hairy root cultures. Plant Cell Tiss Organ Cult 41, 249–258 (1995). https://doi.org/10.1007/BF00045089
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00045089