Producing Friable Callus for Suspension Culture in Glycyrrhiza Glabra

Glycyrrhiza glabra is widely used in bronchial problems such as catarrh, bronchitis, cold, flu and coughs and this plant has a useful secondary metabolite by the name of Lycorys and widely extracted from Bioreactors or hairy roots techniques, the initial step in aim to start a bioreactor is producing a friable callus which use for a suspension culture , A hormonal balance would be vital for producing this kind of callus, An experiment was conducted by the base of NAA and 2,4-D as auxins and 6-BA as cytokinins plant growing regulators in rang of 0.5 2.0 mg/l in a MS medium, Each experiment contained at least 20 replicates and the experiments were repeated three times. Data were documented up to 5 weeks of culture, well grown callus induced from explants were selected to transfer to MS medium with appropriate hormones for subculture. no calli were induced but When 2,4-D was used into MS medium, all explants formed white callus and induction rates increased with the increase of concentration of 2,4-D. It was found that the addition of high concentration of 6-BA into the MS medium in combination with 2,4-D had prompt effect on callus formation ,The maximum induction rate was recorded as 96 % in MS medium with 0.5 mg/l 2,4-D and 2 mg/l 6-BA with a light compact structure . However, MS medium with single 6-BA induced no callus in none of explants, although the combination of NAA and 6-BA could induce callus formation in explants, but callus turned brown. As previous primary experiments indicated that only high concentration of BA in combination with 2, 4-D or NAA in medium was relatively more suitable for subculture, the well grown callus were Selected for subculture on MS medium with 2.0 mg/l 6-BA in combination with 0.5 2 mg/l 2, 4-D or with 0.5 2.0 mg/l NAA to test their growing state. 40 days later after the callus were subculture on the MS with 6-BA and 2,4-D, they turned from green to white and became compact, while callus cultured on MS medium with 6BA and NAA became loose, and spongy which was more suitable for continuous subculture and suspension culture. Therefore, MS medium supplemented with 0.5 2.0 mg/l NAA and 2.0 mg/l 6-BA was used for subsequent experiments. Through testing, it was found that the MS medium containing 2 mg/l 6-BA and 0.5 mg/l NAA had the maximum effect and was decided for subculture and cell suspension culture , By the base of this experiment and due the producing a friable callus for our aim its seems that explants of Glycyrrhiza glabra on MS medium by the maximum induction rate was recorded as 96 % in MS medium with 0.5 mg/l 2,4-D and 2 mg/l 6-BA with a light compact structure but it’s not a suitable for callus induction ,using a hormonal dense with 1.0 mg/l NAA and 2 mg/l 6-BA by 91.6 induction rate with a Loose, spongy, friable structure which use in a suspension culture. Key word: Glycyrrhiza glabra, suspension culture, callus inducing 126 Adv. Environ. Biol., 3(2): 125-128, 2009