Plant regeneration and production of embelin from organogenic and embryogenic callus cultures of Embelia ribes Burm. f.-a vulnerable medicinal plant

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Journal Article
In Vitro Cellular and Developmental Biology - Plant, 2011, 47 (4), pp. 506 - 515
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Embelia ribes, an important vulnerable medicinal liana, was regenerated through organogenesis and embryogenesis using leaf explants. Leaf explants produced organogenic calluses on MS medium supplemented with 1.0 mg l-12,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 mg l-16-benzylaminopurine. Shoot regeneration was obtained from organogenic calluses on MS medium containing different concentrations of thidiazuron (TDZ) and indole-3-acetic acid (IAA). The frequency of shoot bud organogenesis was highest (23.9 shoots/explant) in MS medium containing 0. 5 mg l-1TDZ and 0.1 mg l-1IAA. The best result for induction of embryogenic callus was noticed in the combination of 2.0 mg l-1TDZ and 0.5 mg l-12,4-D. This callus, maintained in the same medium, showed the highest differentiation of embryos (56.5%) after 6 wk of culture. Embryos were transferred to MS medium supplemented with different concentrations of TDZ, and this facilitated conversion of embryos into plants. After 6 wk of subculture, MS medium with 0. 05 mg l-1TDZ favored the highest percentage (52.2%) embryo conversion. As per the present protocol, 52.2% of the embryos underwent conversion, and a mean number of 29.5 shoots per culture was obtained. Shoots developed from both types of calluses were rooted on half-strength MS basal medium supplemented with 1.0 mg l-1indole-3-butyric acid. HPLC-UV assay demonstrated the highest embelin content (5.33% w/w) in the embryogenic callus cultures. Embelin was isolated from embryogenic callus and was identified using IR and1H NMR studies. © 2011 The Society for In Vitro Biology.
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