Abstract
Withanolide is one of the most extensively exploited steroidal lactones, which are biosynthesized in Withania somnifera. Its production from cell suspension culture was analyzed to defeat limitations coupled with its regular supply from the plant organs. In order to optimize the different factors for sustainable production of withanolides and biomass accumulations, different concentrations of auxins or cytokinins and their combinations, carbon sources, agitation speed, organic additives and seaweed extracts was studied in cell suspension culture. Maximum biomass accumulation (16.72 g fresh weight [FW] and 4.18 g dry weight [DW]) and withanolides production (withanolide A 7.21 mg/g DW, withanolide B 4.23 mg/g DW, withaferin A 3.88 mg/g DW and withanone 6.72 mg/g DW) were achieved in the treatment of Gracilaria edulis extract at 40 % level. Organic additive l-glutamine at 200 mg/l in combination with picloram (1 mg/l) and KN (0.5 mg/l) promoted growth characteristics (11.87 g FW and 2.96 g DW) and withanolides synthesis (withanolide A 5.04 mg/g DW, withanolide B 2.59 mg/g DW, withaferin A 2.36 mg/g DW and withanone 4.32 mg/g DW). Sucrose at 5 % level revolved out to be a superior carbon source yielded highest withanolides production (withanolide A 2.88 mg/g DW, withanolide B 1.48 mg/g DW, withaferin A 1.35 mg/g DW and withanone 2.47 mg/g DW), whereas biomass (7.28 g FW and 1.82 g DW) was gratefully increased at 2 % level of sucrose in cell suspension culture. This optimized protocol can be utilized for large scale cultivation of W. somnifera cells in industrial bioreactors for mass synthesis of major withanolides.
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Abbreviations
- 2,4-D:
-
2,4-Dichlorophenoxy acetic acid
- 2iP:
-
6-(α,α-Dimethylallylamino)-purine
- BA:
-
6-Benzyladenine
- DW:
-
Dry weight
- FW:
-
Fresh weight
- HPLC:
-
High-performance liquid chromatography
- IAA:
-
Indole-3-acetic acid
- IBA:
-
Indole-3-butyric acid
- KN:
-
Kinetin
- MS:
-
Murashige and Skoog medium
- NAA:
-
α-Naphthaleneacetic acid
- PGRs:
-
Plant growth regulators
- ROS:
-
Reactive oxygen species
- rpm:
-
Rotation per minute
- SWEs:
-
Seaweed extracts
- SWs:
-
Seaweeds
- SWE:
-
Seaweed extract
- TDZ:
-
Thiadiazuron
- Zea:
-
Zeatin
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Acknowledgments
Prof. A. Ganapathi is thankful to the University Grants Commissions (UGC), Government of India, for the award of UGC-BSR (Basic Scientific Research Fellowship). Dr. M. Manickavasagam is grateful to the University Grants Commissions (UGC), Government of India, for providing UGC-STRAT UP Grant. The first author thankfully acknowledges the Council of Scientific and Industrial Research (CSIR), Government of India, for the award of CSIR-SRF. G. Kapil Dev, M. Jeyaraj and M. Rajesh gratefully acknowledge the Bharathidasan University, UGC and CSIR, Govt. of India for the award of University Research Fellow (BDU-URF), Rajivi Gandhi National Fellowship (UGC RGNF) and Senior Research Fellow (SRF), respectiviely. All authors acknowledge the CSIR, Government of India, for providing financial support to carry out this work. The authors are thankful to Dr. R. Boopathy, head of the department, and Dr. S. Girija of the Department of Biotechnology, Bharathiar University, Coimbatore, for providing the HPLC facility.
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Sivanandhan, G., Kapil Dev, G., Jeyaraj, M. et al. A promising approach on biomass accumulation and withanolides production in cell suspension culture of Withania somnifera (L.) Dunal. Protoplasma 250, 885–898 (2013). https://doi.org/10.1007/s00709-012-0471-x
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DOI: https://doi.org/10.1007/s00709-012-0471-x