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Comprehensive assessment of the genes involved in withanolide biosynthesis from Withania somnifera: chemotype-specific and elicitor-responsive expression

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Abstract

Withania somnifera (L.) Dunal (Family, Solanaceae), is among the most valuable medicinal plants used in Ayurveda owing to its rich reservoir of pharmaceutically active secondary metabolites known as withanolides. Withanolides are C28-steroidal lactones having a triterpenoidal metabolic origin synthesised via mevalonate (MVA) pathway and methyl-D-erythritol-4-phosphate (MEP) pathway involving metabolic intermediacy of 24-methylene (C30-terpenoid) cholesterol. Phytochemical studies suggest differences in the content and/or nature of withanolides in different tissues of different chemotypes. Though development of genomic resources has provided information about putative genes encoding enzymes for biosynthesis of intermediate steps of terpenoid backbone, not much is known about their regulation and response to elicitation. In this study, we generated detailed molecular information about genes catalysing key regulatory steps of withanolide biosynthetic pathway. The full-length sequences of genes encoding enzymes for intermediate steps of terpenoid backbone biosynthesis and their paralogs have been characterized for their functional and structural properties as well as phylogeny using bioinformatics approach. The expression analysis suggests that these genes are differentially expressed in different tissues (with maximal expression in young leaf), chemotypes and in response to salicylic acid (SA) and methyl jasmonate (MJ) treatments. Sub-cellular localization studies suggest that both paralogs of sterol ∆-7 reductase (WsDWF5-1 and WsDWF5-2) are localized in the endoplasmic reticulum (ER) thus supporting their indispensible role in withanolide biosynthesis. Comprehensive information developed, in this study, will lead to elucidation of chemotype- as well as tissue-specific withanolide biosynthesis and development of new tools for functional genomics in this important medicinal plant.

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Acknowledgements

The authors are thankful to the Council of Scientific and Industrial Research, New Delhi, Govt. of India, for providing the financial support to carry out this work under NMITLI scheme. AVA, PG and DS acknowledge Indian Council of Medical Research (ICMR), Council of Scientific and Industrial Research (CSIR) and University Grants Commission (UGC) for Junior and Senior Research Fellowships. The authors acknowledge Dr. Neelam Singh Sangwan and Dr. Rajender Singh Sangwan, CSIR-Central Institute of Medicinal and Aromatic Plants (CSIR-CIMAP), Lucknow, for developing and sharing Withania chemotypes under NMITLI scheme. The authors acknowledge Dr. Surendra Pratap Singh, CSIR-NBRI for help in confocal microscopy.

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  1. Parul Gupta

    Present address: Botany and Plant Pathology, Oregon State University, 2082-Cordley Hall, Corvallis, OR, 97331, USA

Authors and Affiliations

  1. Council of Scientific and Industrial Research, National Botanical Research Institute, Rana Pratap Marg, Lucknow, 226001, India

    Aditya Vikram Agarwal, Parul Gupta, Deeksha Singh, Yogeshwar Vikram Dhar & Prabodh Kumar Trivedi

  2. Department of Biochemistry, University of Lucknow, Lucknow, 226007, India

    Aditya Vikram Agarwal & Deepak Chandra

  3. Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, 2 Rafi Marg, New Delhi, 110001, India

    Deeksha Singh, Yogeshwar Vikram Dhar & Prabodh Kumar Trivedi

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  1. Aditya Vikram Agarwal
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Correspondence to Prabodh Kumar Trivedi.

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Agarwal, A.V., Gupta, P., Singh, D. et al. Comprehensive assessment of the genes involved in withanolide biosynthesis from Withania somnifera: chemotype-specific and elicitor-responsive expression. Funct Integr Genomics 17, 477–490 (2017). https://doi.org/10.1007/s10142-017-0548-x

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