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

  • Plant Tissue Culture
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Abstract

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.

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Acknowledgments

Authors gratefully acknowledge Dr. P. S. Udayan, Senior Scientist, Botany Division, Centre for Medicinal Plants Research for collection and identifying the plant species. The help rendered by Dr. S. Ravi, Senior Scientist, Phytochemistry division, Centre for Medicinal Plants Research, for spectral analysis of the compound is also acknowledged. We are also thankful to Hashim K. M., Phytochemistry division, Centre for Medicinal Plants Research for helpful discussions and comments. Authors are also thankful to the authorities of Arya Vaidya Sala (AVS), Kottakkal for providing facilities and Sir Dorabji Tata Trust, Mumbai for financial support.

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  1. Centre for Medicinal Plants Research (CMPR), Arya Vaidya Sala (AVS), Kottakkal, Malappuram, 676 503, Kerala, India

    A. V. Raghu, Kuzhiyumparambil Unnikrishnan, S. P. Geetha, Gerald Martin & Indira Balachandran

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  1. A. V. Raghu
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  2. Kuzhiyumparambil Unnikrishnan
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  3. S. P. Geetha
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  4. Gerald Martin
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  5. Indira Balachandran
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Correspondence to A. V. Raghu.

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Editor: J. Canhoto

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Raghu, A.V., Unnikrishnan, K., Geetha, S.P. et al. Plant regeneration and production of embelin from organogenic and embryogenic callus cultures of Embelia ribes Burm. f.—a vulnerable medicinal plant. In Vitro Cell.Dev.Biol.-Plant 47, 506–515 (2011). https://doi.org/10.1007/s11627-011-9365-4

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