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Overexpression of a Catharanthus tryptophan decarboxylase (tdc) gene leads to enhanced terpenoid indole alkaloid (TIA) production in transgenic hairy root lines of Rauwolfia serpentina

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Abstract

To enhance the production of terpenoid indole alkaloids in Rauwolfia serpentina, Catharanthus tryptophan decarboxylase (Crtdc) gene was over-expressed in transgenic hairy root cultures using Agrobacterium rhizogenes-mediated transformation. Among six transgenic hairy root lines, line RT4 accumulated the highest alkaloid content, with 0.1202 % dry weight (DW) reserpine and 0.0064 % DW ajmalicine, after 10 weeks of culture. Whereas, wild-type roots accumulated 0.0596 ± 0.003 % DW reserpine and 0.0011 ± 0.001 % DW ajmalicine. Transgenic hairy root line RT7 produced the lowest alkaloid content (reserpine: 0.0896 ± 0.002 % DW; ajmalicine: 0.002 ± 0.0 % DW). On the basis of alkaloid content the six hairy root lines were grouped as RT4/RT2 > RT3/RT5 > RT7/RT8. Analysis of gene expression profile indicated that Crtdc was expressed at a higher level in transgenic lines, which could be correlated with enhanced metabolite accumulation in roots. This study confirms that over-expression of Crtdc is a superlative method to improve the biosynthetic potential of Rauwolfia hairy root cultures. Enhanced reserpine and ajmalicine production can serve as an alternative choice to provide resources for relative pharmaceutical industries.

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Acknowledgments

The authors gratefully acknowledge Director, CIMAP (CSIR) for providing the facility to carry out this work. Financial support rendered by Department of Science and Technology (DST), New Delhi to SM and Council of Scientific and Industrial Research (CSIR), New Delhi to VS is gratefully acknowledged. Thanks are also due to Mr. Praveen Prakash SRF, Plant Biotechnology Division CIMAP, Lucknow for his generous help.

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Author notes

  1. Vikas Srivastava

    Present address: National Institute of Plant Genome Research, Aruna Asaf Ali Road, P.O. Box 10531, New Delhi, 110 067, India

Authors and Affiliations

  1. Plant Biotechnology Division, Central Institute of Medicinal and Aromatic Plants, Picnic Spot Road, P.O. CIMAP, Lucknow, 226015, India

    Shakti Mehrotra, Vikas Srivastava, Laiq Ur Rahman & A. K. Kukreja

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  1. Shakti Mehrotra
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  2. Vikas Srivastava
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  3. Laiq Ur Rahman
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  4. A. K. Kukreja
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Correspondence to Shakti Mehrotra.

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11240_2013_369_MOESM1_ESM.doc

Major steps in terpenoid indole alkaloid biosynthetic pathway. Shaded area in box represents the steps common in Catharanthus and Rauwolfia. AS: nthranilate synthase; TDC: tryptothan decarboxylase; STR: strictosidine synthase etc. (Dashed arrows indicate more than one step) (DOC 243 kb)

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Mehrotra, S., Srivastava, V., Rahman, L.U. et al. Overexpression of a Catharanthus tryptophan decarboxylase (tdc) gene leads to enhanced terpenoid indole alkaloid (TIA) production in transgenic hairy root lines of Rauwolfia serpentina . Plant Cell Tiss Organ Cult 115, 377–384 (2013). https://doi.org/10.1007/s11240-013-0369-0

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  • DOI: https://doi.org/10.1007/s11240-013-0369-0

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