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DNA Barcodes Distinguish Withania somnifera and Withania ashwagandha

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

On the basis of morphology, chemical profiling, crossing ability, amplified fragment length polymorphism and comparison of nucleotide sequences of nuclear ribosomal internal transcribed spacer (ITS), the cultivated form of Withania somnifera, a species of therapeutic value, has been circumscribed as a new species, Withania ashwagandha. The present study was undertaken to ascertain whether the two species can be distinguished on the basis of DNA barcoding. Six barcode loci, ITS, ITS2, matK (maturase K), rbcL (ribulose-bisphosphate carboxylase/oxygenase, large subunit), rpoC1 (RNA polymerase-β′ subunit, main catalytic subunit) and trnH-psbA spacer (transfer RNA for histidine-photosystem II protein D1 spacer) from W. somnifera, W. ashwagandha, their hybrid, and ‘ashwagandha’ market samples were amplified, sequenced, and compared. ITS, ITS2 and matK distinguished two species on the basis of phylogenetic tree method. Likewise, BLAST 1 analysis based on ITS, ITS2, matK, and rbcL individually discriminated two species. However, on the basis of Kimura 2 Parameter distances, two species could not be distinguished as the requirement of a distinct barcode gap—the highest intraspecific distance being lower than the lowest interspecific distance—was not met by any of the loci. If compared by character-based method, ITS, ITS2 and matK sequences of the two species had distinct diagnostic nucleotides (pure character attributes) at nine, four and one positions, respectively. Interestingly, all market samples co-segregated and shared character attributes with W. ashwagandha.

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Acknowledgements

The authors thank Dr. Sushma Koul formerly from the Biodiversity and Applied Botany Division, Institute of Integrative Medicine, Jammu for providing the plant material. They gratefully acknowledge the financial support provided by the Indian Council of Medical Research, New Delhi to S.B.B. in form of a research project. S.M. too thankfully acknowledges the receipt of Junior and Senior Research fellowships from this project. The present work was also partially supported by a Research and Development grant provided to S.B.B. by the University of Delhi. The language editing of the manuscript by Dr. Anupam Raina, a professional copy editor, is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Botany, University of Delhi, Delhi, 110007, India

    Saloni Malik, Bilal A. Mir, Hemant K. Singh & Shashi B. Babbar

  2. Department of Botany, School of Life Sciences, Satellite Campus Kargil, University of Kashmir, Kargil, Jammu and Kashmir, 194103, India

    Bilal A. Mir

  3. Amity Institute of Biotechnology, Amity University, Sector-125, Noida, UP, 201303, India

    Manju Chaudhary & Soom N. Raina

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  1. Saloni Malik
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Correspondence to Shashi B. Babbar.

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Significance statement

Withania somnifera, ‘Ashwagandha’, is an important medicinal plant. Because of its high demand, it has been domesticated and is cultivated. There have many suggestions that its cultivated form has diverged enough from the parent species to be treated as a new species, W. ashwagandha. The present work, based on DNA barcoding, confirms its species-level status and provides an effective identification tool for checking substitutions in its market samples.

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Malik, S., Mir, B.A., Singh, H.K. et al. DNA Barcodes Distinguish Withania somnifera and Withania ashwagandha . Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 88, 1413–1424 (2018). https://doi.org/10.1007/s40011-017-0879-3

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  • DOI: https://doi.org/10.1007/s40011-017-0879-3

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