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Influence of cellular differentiation and elicitation on intermediate and late steps of terpenoid indole alkaloid biosynthesis in Catharanthus roseus

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Abstract

The invaluable antineoplastic bisindole alkaloids of Catharanthus roseus and their precursor, vindoline, are not produced in cell cultures. The intricacies involved in endogenous (cellular differentiation) and exogenous (elicitation) regulation of their biosynthesis need to be dissected out for favorable exploitation. This study aimed at elucidating the effect of Pythium aphanidermatum homogenate and methyl jasmonate (MeJa) on in vitro cultures (of cv. ‘Dhawal’) representing increasing level of differentiation (suspension < callus < shoots) in terms of alkaloid accumulation and transcript abundance of strictosidine β-d-glucosidase (SGD) and acetyl-CoA: 4-O-deacetylvindoline 4-O-acetyl-transferase (DAT) genes, representing intermediate and late steps, respectively, of terpenoid indole alkaloid biosynthesis. Elicitation of suspension cultures caused transcriptional upregulation of SGD and enhanced the accumulation of total alkaloids but did not produce vindoline as DAT transcripts were always found to be absent in suspension-cultured cells. Vindoline was also not detected in unelicited and MeJa-treated callus but appeared upon elicitation with fungal homogenate for 24 h that coincided with maximal DAT transcription. Transcript levels of both genes increased upon elicitation of callus but remained below levels present in the mature plant leaf. Elicitation caused appearance of vindoline in shoots and increased the transcript abundance of both genes beyond levels observed in the mature plant leaf. Differentiation was essential for expression of DAT but not SGD, and vindoline biosynthetic potential increased with it.

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Acknowledgments

We are grateful to the Council of Scientific and Industrial Research (CSIR), India, and the Department of Biotechnology, India, for the financial support extended by them. We also thank Dr. R.N. Kulkarni, Senior Scientist, CIMAP Resource Centre, Bangalore, for providing cv. ‘Dhawal’ seeds.

Conflict of interest

The authors declare that they have no conflict of interest.

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  1. Suman P. S. Khanuja

    Present address: C41-42, Double Storey, Ramesh Nagar, New Delhi, 110015, India

Authors and Affiliations

  1. Genetic Resources and Biotechnology Division, Central Institute of Medicinal and Aromatic Plants (CSIR), P.O. CIMAP, Lucknow, 226015, India

    Ashutosh K. Shukla, Ajit K. Shasany & Suman P. S. Khanuja

  2. Analytical Chemistry Division, Central Institute of Medicinal and Aromatic Plants (CSIR), P.O. CIMAP, Lucknow, 226015, India

    Ram K. Verma & Madan M. Gupta

  3. Plant Tissue Culture Division, Central Institute of Medicinal and Aromatic Plants (CSIR), P.O. CIMAP, Lucknow, 226015, India

    Ajay K. Mathur

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  1. Ashutosh K. Shukla
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Correspondence to Suman P. S. Khanuja.

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Shukla, A.K., Shasany, A.K., Verma, R.K. et al. Influence of cellular differentiation and elicitation on intermediate and late steps of terpenoid indole alkaloid biosynthesis in Catharanthus roseus . Protoplasma 242, 35–47 (2010). https://doi.org/10.1007/s00709-010-0120-1

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