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

• Published in: In vitro cellular & developmental biology. Plant 2011-08, Vol.47 (4), p.506-515
• Main Authors: Raghu, A. V, Unnikrishnan, Kuzhiyumparambil, Geetha, S. P, Martin, Gerald, Balachandran, Indira
• Format: Article
• Language: English
• Subjects: 2,4-D; Acetic acid; Acids; Ayurvedic medicine; Berries; Biomedical and Life Sciences; Callus; Cell Biology; Developmental Biology; Embelia ribes; embryo (plant); Embryogenesis; Embryonic growth stage; Embryos; Flowers & plants; indole acetic acid; indole butyric acid; Leaves; Life Sciences; Liquid chromatography; Medicinal plants; Metabolites; MICROPROPAGATION; nuclear magnetic resonance spectroscopy; Organogenesis; Plant Breeding/Biotechnology; Plant cells; Plant Genetics and Genomics; Plant Sciences; Plant Tissue Culture; Plants; Shoots; Somatic embryogenesis; Somatic embryos; Studies; Sucrose
• ISSN: 1054-5476; 1475-2689
• DOI: 10.1007/s11627-011-9365-4

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−1 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 mg l−1 6-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−1 TDZ and 0.1 mg l−1 IAA. The best result for induction of embryogenic callus was noticed in the combination of 2.0 mg l−1 TDZ and 0.5 mg l−1 2,4-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−1 TDZ 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−1 indole-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 and 1H NMR studies.