Direct rhizogenesis from leaf explants and establishment of an in vitro stolon culture system and subsequent plant regeneration for Glycyrrhiza glabra have been described. MS liquid medium supplemented with 0.01 mg l-1 of NAA was most effective for stolon proliferation. Extensive proliferation of stolon and shoot regeneration was achieved on medium containing 3 % sucrose with 0.01 mg l-1 NAA. Stolons with nodes showing growth was transferred under light for plantlet regeneration in the same medium. This paper is the first report in G. glabra describing a complete regeneration procedure via in vitro stolon proliferation along with quantitative data for glycyrrhizin and genetic fidelity of plant regenerated in vitro there from. In vitro stolon proliferation described here would be an efficient way for regeneration of plants for functional genomics studies and better understanding of glycyrrhizin (GA) metabolism.
Medicinal Chemistry Division, CSIR-Indian Institute of Integrative Medicine, Jammu 180 001, India
Bibliografia
Anonymous (2005) A dictionary of Indian raw materials and industrial products, first supplement series. In: The wealth of India. National Institute of Sciences Communication and Information Resources, CSIR, New Delhi, 3, pp 195–198
Ayabe S, Takano H, Fujita T, Furuya T, Hirota H, Takahashi T (1990) Triterpenoid biosynthesis in tissue cultures of Glycyrrhiza glabra var. glandulifera. Plant Cell Rep 9:181–184
Cinatl J, Morgenstern B, Bauer G, Chandra P, Rabenau H, Doerr HW (2003) Glycyrrhizin, an active component of liquorice roots, and replication of SARS-associated coronavirus. Lancet 361:2045–2046
Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15
Finney RSH, Somers GF (1958) The antiinflammatory activity of glycyrrhetinic acid and derivatives. J PharmPharmacol 10:613–620
Fiore C, Eisenhut M, Krausse R, Ragazzi E, Pellati D, Armanini D, Bielenberg J (2008) Antiviral effects of Glycyrrhiza species. Phytother Res 22:141–148
Franklin G, Sheeba CJ, Laksmisita G (2004) Regeneration of eggplant (Solanum melongena L.) from root explants. In Vitro Cell Dev Biol Plant 40:188–191
Gupta V, Kak A, Singh BB (1997) Studies on seed germination and seedling vigour in liquorice (Glycyrrhiza glabra). JMAPS 2:412–413
Hayashi H, Fukui H, Tabata M (1988) Examination of triterpenoids produced by callus and cell suspension cultures of Glycyrrhiza glabra. Plant Cell Rep 7:508–511
Henry M, Edy MA, Desmarest P, Manoir DJ (1991) Glycyrrhiza glabra L. (Licorice): cell culture, regeneration, and the production of glycyrrhizin. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, vol 15, Medicinal and aromatic plants III. Springer, Berlin, pp 270–282
Ito M, Sato A, Hirabayashi K, Tanabe F, Shigeta S, Baba M, Clercq ED, Nakashima H, Yamamoto N (1988) Mechanism of inhibitory effect of glycyrrhizin on replication of human immunodeficiency virus (HIV). Antiviral Res 10:289–298
Jatav VS, Singh SK, Khatri P, Sharma AK (2011) Recent pharmacological trends of Glycyrrhiza glabra Linn. Unani Res 1:1–4
Kojoma M, Ohyama K, Seki H, Hiraoka Y, Asazu SN, Sawa S, Sekizaki H, Yoshida S, Muranaka T (2010) In vitro proliferation and triterpenoid characteristics of licorice (Glycyrrhiza uralensis Fischer, Leguminosae) stolons. Plant Biotechnol 27:59–66
Li W, Asad Y, Yoshikawa T (2000) Flavonoid constituents from Glycyrrhiza glabra hairy root cultures. Phytochemistry 55:447–456
Mehrotra S, Kukreja AK, Khanuja SPS, Mishra BN (2008) Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactor. Electron J Biotechnol 11:1–7
Mehrotra S, Khwaja O, Kukreja AK, Rahman L (2012) ISSR and RAPD based evaluation of genetic stability of encapsulated micro shoots of Glycyrrhiza glabra following six months of storage. Mol Biotechnol 52:262–268
Mousa N, Siagura P, Wiryowidagdo S, Wagih ME (2006) Rapid clonal propagation of licorice (Glycyrrhiza glabra) by in vitro shoot culture. Sugar Tech 8:292–298
Nagat A, Mousa TR, Siaguru I, Wiryowidagdo S, Wagih ME (2007) Establishment of regenerative callus and cell suspension system of licorice (Glycyrrhiza glabra) for the production of the sweetener glycyrrhizin in vitro. Sugar Tech 9:72–82
Park HY, Park SH, Yoon HK, Han MJ, Kim DH (2004) Anti-allergic activity of 18b-glycyrrhetinic acid-3-O-beta-D glucuronide. Arch Pharm Res 27:57–60
Saito K, Kaneko H, Yamazaki M, Yoshida M, Murakoshi I (1990) Stable transfer and expression of chimeric genes in licorice (Glycyrrhiza uralensis) using a Ri plasmid binary vector. Plant Cell Rep 8:718–721
Shah RR, Dalal KC (1980) In vitro multiplication of Glycyrrhiza. Curr Sci 49:69–71
Shirazi Z, Piri K, Asl AM, Hasanloo T (2012) Glycyrrhizin and isoliquiritigenin production by hairy root culture of Glycyrrhiza glabra. J Med Plants Res 6:4640–4646
Thengane R, Kulkarni DK, Krishnamurthy KV (1998) Micropropagation of licorice (Glycyrrhiza glabra l.) through shoot tip and nodal cultures. In Vitro CeIl Dev Biol Plant 34:331–334
Toivonen L, Rosenqvist H (1995) Establishment and growth characteristics of Glycyrrhiza glabra hairy root cultures. Plant Cell Tiss Org Cult 41:249–258
Wongwicha W, Tanaka H, Shoyama Y, Tuvshintogtokh I, Putalun W (2008) Production of glycyrrhizin in callus cultures of licorice. Z Naturforsch 63:413–417
Yoo S, Kim S (1976) Studies in tissue culture of medicinal plants (II). Tissue culture of Glycyrrhiza glabra L. var. glandulifera Reg. et Herder. Kor J Pharmacol 7:55D57
Zhang MZ, Jie X, Bing Y, Huiyong Y, Qiuyin C, Martha J, Shrubsole XC, Valentina K, Wei Z, Ambra P, Harris RC (2009) Inhibition of 11b-hydroxysteroid dehydrogenase type II selectively blocks the tumor COX-2 pathway and suppresses colon carcinogenesis in mice and humans. J Clin Invest 119:876–885
Zietkiewicz E, Rafalski A, Labuda D (1994) Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20:176–183