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Effect of culture conditions, cytokinins, methyl jasmonate and salicylic acid on the biomass accumulation and production of withanolides in multiple shoot culture of Withania somnifera (L.) Dunal using liquid culture

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Abstract

The influence of cytokinins and culture conditions including medium volume, harvest time and elicitation with abiotic elicitors (SA/MeJ) have been studied for the optimal production of biomass and withanolides in the multiple shoot culture of Withania somnifera. Elicitation of shoot inoculum mass (2 g l−l FW) with SA at 100 μM in the presence of 0.6 mg l−l BA and 20 mg l−l spermidine for 4 h exposure time at the 4th week in 20 ml liquid medium recorded higher withanolides production (withanolides A [8.48 mg g−l DW], withanolides B [15.47 mg g−l DW], withaferin A [29.55 mg g−l DW] and withanone [23.44 mg g−l DW]), which were 1.14 to 1.18-fold higher than elicitation with MeJ at 100 μM after 5 weeks of culture. SA-elicited cultures did not exhibit much variation in biomass accumulation when compared to control. This cytokinin induces and SA-elicited multiple shoot culture protocol provides a potential alternative for the optimal production of biomass and withanolides utilizing liquid culture.

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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

JA:

Jasmonic acid

KN:

Kinetin

MeJ:

Methyl jasmonate

MS:

Murashige and Skoog medium

NN:

Nitsch and Nitsch medium

PGR:

Plant growth regulators

RT:

Retention time

SA:

Salicylic acid

TDZ:

Thiadiazuron

Zea:

Zeatin

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Acknowledgments

The first author gratefully acknowledges the Council of Scientific and Industrial Research (CSIR), Government of India, for the award of CSIR-SRF. All authors acknowledge CSIR, Government of India, for providing financial support to carry out this work. The authors thankful to Dr. S. Girija, Associate Professor, Department of Biotechnology, Bharathiar University, Coimbatore for providing HPLC facility.

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Authors and Affiliations

  1. Department of Biotechnology and Genetic Engineering, School of Biotechnology, Bharathidasan University, Tiruchirappalli, 620 024, Tamil Nadu, India

    Ganeshan Sivanandhan, Manoharan Rajesh, Muthukrishnan Arun, Murugaraj Jeyaraj, Gnanajothi Kapil Dev, Annavi Arjunan, Markandan Manickavasagam, Manickam Muthuselvam & Andy Ganapathi

  2. Periyar E. V. R College (Autonomous), Tiruchirappalli, 620 023, Tamil Nadu, India

    Natesan Selvaraj

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  1. Ganeshan Sivanandhan
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  10. Andy Ganapathi
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Correspondence to Andy Ganapathi.

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Communicated by J. Ueda.

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Sivanandhan, G., Rajesh, M., Arun, M. et al. Effect of culture conditions, cytokinins, methyl jasmonate and salicylic acid on the biomass accumulation and production of withanolides in multiple shoot culture of Withania somnifera (L.) Dunal using liquid culture. Acta Physiol Plant 35, 715–728 (2013). https://doi.org/10.1007/s11738-012-1112-x

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  • DOI: https://doi.org/10.1007/s11738-012-1112-x

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