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Factors influencing podophyllotoxin production in adventitious root culture of Podophyllum hexandrum Royle

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Abstract

To determine the possibility of generating the Podophyllotoxin accumulation using Podophyllum hexandrum adventitious roots derived from root segments, several nutrient constituents (carbon sources, media strength, initial medium pH, ammonium and nitrate proportion and phosphate ratio) were evaluated in culture. The maximum biomass accumulation was recorded in 0.50 MS medium containing 3 mg/l indole-3-butyric acid and 2 % sucrose, and the maximum accumulation of Podophyllotoxin was documented in the same strength of MS medium with 6 % of sucrose. When the initial medium pH was on 6 in the optimized MS medium, the biomass and Podophyllotoxin accumulations were highest. The lower concentration of ammonium (10 mM) in combination with a moderate concentration of nitrate (20 mM) was found ideal for maximum accumulations of biomass and Podophyllotoxin. Maximum Podophyllotoxin accumulation (6.4 mg/g dry weight) was recorded at the higher concentration of phosphate (2.25 mM), and lower concentration of phosphate (1.25 mM) showed highest growth accumulation. The outcome of the present work will be helpful for the large-scale cultivation of adventitious root for the production of Podophyllotoxin.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxy acetic acid

DW:

Dry weight

FW:

Fresh weight

HPLC:

High performance liquid chromatography

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

MS:

Murashige and Skoog medium

NAA:

1-Naphthaleneacetic acid

NH4 +/NO3 − :

Ammonium/nitrate ratio

PGRs:

Plant growth regulators

PTOX:

Podophyllotoxin

RT:

Retention time

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Acknowledgments

Prof. A. Ganapathi is thankful to University Grants Commissions (UGC), Government of India, for the award of UGC-BSR (Basic Scientific Research Fellowship). Dr. M. Manickavasagam is grateful to University Grants Commissions (UGC), Government of India, for providing UGC-START UP Grant. First author is thankful to Prof. S. K. Nandi, G. B. Pant Institute, Almora, Uttaranchal and Dr. Lokho Puni IFS, Forest Research Institute, Dehradun, Uttaranchal for helping in collection of seeds. The corresponding author is thankful to DRDO, Government of India, for providing financial support (DLS/81/48222/LSRB–171 BTB/2008) to carry out this work.

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Authors and Affiliations

  1. Department of Biotechnology and Genetic Engineering, School of Biotechnology, Bharathidasan University, Tiruchirappalli, 620024, Tamil Nadu, India

    Manoharan Rajesh, Ganeshan Sivanandhan, Venkatachalam Vasudevan, Jeevaraj Theboral, Markandan Manickavasagam & Andy Ganapathi

  2. Department of Genetic Engineering, School of Bioengineering, SRM University, Kattankulathur, Kancheepuram, 603203, Tamil Nadu, India

    Muthukrishnan Arun

  3. Department of Biotechnology, Bharathiar University, Coimbatore, Tamil Nadu, India

    Shanmugam Girija

  4. Government Arts College, Kulithalai, 639120, Tamil Nadu, India

    Natesan Selvaraj

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  1. Manoharan Rajesh
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  2. Ganeshan Sivanandhan
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  3. Muthukrishnan Arun
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  4. Venkatachalam Vasudevan
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  7. Markandan Manickavasagam
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  9. Andy Ganapathi
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Correspondence to Andy Ganapathi.

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Communicated by K.-Y. Paek.

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Rajesh, M., Sivanandhan, G., Arun, M. et al. Factors influencing podophyllotoxin production in adventitious root culture of Podophyllum hexandrum Royle. Acta Physiol Plant 36, 1009–1021 (2014). https://doi.org/10.1007/s11738-013-1479-3

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  • DOI: https://doi.org/10.1007/s11738-013-1479-3

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