Title: Producing Friable Callus for Suspension Culture in Glycyrrhiza Glabra
Abstract: Glycyrrhiza glabra is widely used in bronchial problems such as catarrh, bronchitis, cold, flu and coughs. This plant contains a valuable secondary metabolite by the name of Lycorys, which could be extracted from hairy roots of G. glabra. Nevertheless Lycorys could also be isolated from suspension culture of G.glabra in bioreactor. The initial step for culturing G. glabra is to produce friable callus which can be used for suspension culture. A hormonal balance would be vital for induction of this kind of callus, thus an experiment was conducted based on using NAA and 2,4-D as auxins and 6-BA as cytokinins in a rang from 0.5 to 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 all explants formed white callus and induction rates increased with the increase of concentration of 2,4-D. It was found out 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 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 into brown. As previous primary experiments indicated, only high concentrations of BA in combination with 2, 4-D or NAA in medium were 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 6- BA 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 out 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. Based on our experiment 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, nevertheless 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 was the best option for induction of callus for suspension culture.
Publication Year: 2009
Publication Date: 2009-02-01
Language: en
Type: article
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Cited By Count: 1
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