Skip to main content
SpringerLink
Log in

Utility of a multidisciplinary approach for genome diagnostics of cultivated and wild germplasm resources of medicinal Withania somnifera, and the status of new species, W. ashwagandha, in the cultivated taxon

  • Original Article
  • Published:
Plant Systematics and Evolution Aims and scope Submit manuscript

Abstract

Realizing the inconsistencies that exist in the extent and nature of differentiation in the Withania somnifera genetic resources in India, the 21 cultivated and wild accessions, and the two hybrids (cultivated × wild accessions and vice versa) were investigated for morphological, cytogenetical, chemical profiling, and crossability features. Their nuclear and chloroplast genomes were also assayed at the nucleotide sequence level, and by use of DNA markers. Chloroplast DNA diversity and somatic chromosome number (2n = 48) were not helpful in identifying the differences. Other approaches, on the other hand, especially restriction endonuclease digests, types and sequence length composition of ITS 1 and ITS 2 of nuclear ribosomal DNA, AFLP fingerprinting, and crossability barriers unambiguously provided startling discrete differences between the cultivated and wild accessions, indicating a clear division of W. somnifera into two distinct lineages. These data, therefore, are indicative of the fact that because of the unique characteristics of its nuclear genome, and strong crossability barriers vis-à-vis wild accessions of W. somnifera, the cultivated accessions should be relegated to the rank of the separate species, W. ashwagandha.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Aggarwal RK, Brar DS, Nandi S, Huang N, Khush GS (1999) Phylogenetic relationships among Oryza species revealed by AFLP markers. Theor Appl Genet 98:1320–1328

    Article  CAS  Google Scholar 

  • Anon (1976) The wealth of India, raw materials vol. X: SpW. Publications and Information Directorate, CSIR, New Delhi, pp 581–585

  • Archak S, Galkwad AB, Gautam D, Rao EVVB, Swamy KRM, Karihaloo JL (2003) Comparative assessment of DNA fingerprinting techniques (RAPD, ISSR and AFLP) for genetic analysis of cashew (Anacardium occidentale L.) accessions of India. Genome 46:362–369

    Article  CAS  PubMed  Google Scholar 

  • Atal CK, Schwarting AE (1961) Ashwagandha––an ancient Indian drug. Eco Bot 15:256–263

    Article  Google Scholar 

  • Atal CK, Schwarting AE (1962) Intraspecific variability in Withania somnifera. I. A preliminary survey. Llyodia 25:78–87

    Google Scholar 

  • Atal CK, Gupta OP, Raghunathan K, Dhar KL (1975) Pharmacognosy and phytochemistry of Withania somnifera (L.) Dunal (Ashwagandha). Central Council for Research in Indian Medicine & Homeopathy, New Delhi

    Google Scholar 

  • Bhattacharya SK, Goel RK, Kaur R, Ghosal S (1987) Antistress activity of sitoinosides VII and VIII, new acylsteryl glucosides from Withania somnifera. Phytother Res 9:10–13

    Google Scholar 

  • Crawford DJ (1983) Phylogenetic and systematic inferences from electrophoretic studies. In: Tanksley SD, Orton TJ (eds) Isozymes in plant genetics and breeding, part A. Elsevier, Amsterdam, pp 257–287

    Google Scholar 

  • Demesure B, Sodzi N, Petit RJ (1995) A set of universal primers for amplification of polymorphic non-coding regions of mitochondrial and chloroplast DNA in plants. Mol Ecol 4:129–131

    Article  CAS  PubMed  Google Scholar 

  • Dhalla NS, Gupta KC, Sastry NS, Malhotra CL (1961) Comparative studies on Withania somnifera Dunnl. and Withania ashwagandha Kaul. Ind J Pharmacol 23:126–127

    Google Scholar 

  • Goel S, Raina SN, Ogihara Y (2002) Molecular evolution and phylogenetic implications of internal transcribed spacer sequences of nuclear ribosomal DNA in the Phaseolus vigna complex. Mol Phylogenet Evol l22:1–19

    Article  Google Scholar 

  • Gottlieb LD (1981) Electrophoretic evidence and plant populations. In: Reinhold L, Harborne JB, Swain T (eds) Progress in phytochemistry. Pergamon, Oxford, pp 1–46

    Google Scholar 

  • Kaul KN (1957) The origin, distribution and cultivation of Ashwagandha the so called Withania somnifera of Indian Literature. In: Symposium on the Utilization of Indian Medicinal Plants. Council of Scientific & Industrial Research, New Delhi, pp 7–8

  • Kaul MK, Kumar A, Sharma A (2005) Reproductive biology of Withania somnifera (L.) Dunal. Curr Sci 88:1375–1377

    Google Scholar 

  • Khajuria RK, Suri KA, Gupta RK, Satti NK, Amina M, Suri OP, Qazi GN (2004) Separation, Identification and quantification of selected withanolides in plant extracts of Withania somnifera by HPLC–UV (DAD)-positive ion electro spray ionization-mass spectrometry. J Sep Sci 27(7–8):541–546

    Article  CAS  PubMed  Google Scholar 

  • Kothari SK, Singh CP, Kumar YV, Singh K (2003) Morphology, yield and quality of ashwagandha (Withania somnifera L. Dunal) roots and its cultivation economics as influenced by tillage depth and plant population density. J Hort Sci Biotech 78:422–425

    Google Scholar 

  • Kumar A, Kaul MK, Bhan MK, Khanna PK, Suri KA (2007) Morphological and chemical variation in 25 collections of the Indian medicinal plant Withania somnifera (L.) Dunal (Solanaceae). Genet Resour Crop Evol 54:655–660

    Article  CAS  Google Scholar 

  • MacMaster G, Moller M, Hughes M, Edwards TJ, Bellstedt DU (2005) A new species of Streptocarus from Madagascar. Adansonia 27:131–136

    Google Scholar 

  • Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acid Res 8(19):4321–4326

    Article  CAS  PubMed  Google Scholar 

  • Nagpal R, Tanvir Dar H, Raina SN (2008) Molecular systematics of Brassica and allied genera in subtribes Brassicinae, Raphininae, Moricandiinae and Cakilinae (Brassicacae, tribe Brassiceae); the organization and evolution of ribosomal gene families. Bot J Linn Soc 157:545–557

    Article  Google Scholar 

  • Nei M, Li WH (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. Pro Nat Acad Sci USA 76:5269–5273

    Article  CAS  Google Scholar 

  • Perron M, Perry DJ, Andalo C, Bousquet J (2000) Evidence from sequence-tagged-site markers of a recent progenitor-derivative species pair in conifers. Pro Nat Acad Sci USA 97:11331–11336

    Article  CAS  Google Scholar 

  • Raina SN, Yamamoto K, Murakami M (1989) Intraspecific hybridization and its bearing on chromosome evolution in Vicia narbonensis (Fabaceae). Plant Syst Evol 167:201–217

    Article  Google Scholar 

  • Raina SN, Rani V, Kojima T, Ogihara Y, Singh KP, Devarumath RM (2001) RAPD and ISSR fingerprints as useful genetic markers for analysis of genetic of genetic diversity, varietal identification, and phylogenetic relationships in peanut (Arachis hypogea) cultivars and wild species. Genome 44:763–772

    Article  CAS  PubMed  Google Scholar 

  • Rohlf FJ (1998) NTSYS-pc: numerical taxonomy and multivariate analysis system version 2.02K. Applied Biostatistics, New York

    Google Scholar 

  • Sasanuma T, Sehgal D, Sasakuma T, Raina SN (2008) Phylogenetic analysis of Carthamus species based on the nucleotide sequence of the nuclear SACPD gene and chloroplast trnL-trnF IGS region. Genome 51:1–7

    Article  Google Scholar 

  • Sastry MS, Dhalla NS, Malhotra CL (1960) Chemical examination of Withania ashwagandha Kaul. Indian J Pharmacol 22:205

    Google Scholar 

  • Scartezzini P, Antognoni F, Conte L, Maxia A, Troia A, Poli F (2007) Genetic and phytochemical difference between some Indian and Italian plants of Withania somnifera (L.) Dunal. Nat Prod Res 21:923–932

    Article  CAS  PubMed  Google Scholar 

  • Sehgal D, Raina SN (2005) Genotyping safflower (Carthamus tinctorius L.) cultivars by DNA fingerprints. Euphytica 146:67–76

    Article  CAS  Google Scholar 

  • Sehgal D, Raina SN (2008) DNA markers and germplasm resource diagnostics: New perspectives in crop improvement and conservation strategies. In: Arya ID, Arya S (eds) Utilization of biotechnology in plant sciences. Forest Research Institute, Dehradun, pp 39–53

    Google Scholar 

  • Sehgal D, Bhat V, Raina SN (2008a) Advent of diverse DNA markers to decipher genome sequence polymorphism. In: Kirti PB (ed) Handbook of new technologies for genetic improvement of legumes. CRC Press, New York, pp 477–495

    Google Scholar 

  • Sehgal D, Bhat V, Raina SN (2008b) Applicability of DNA markers for genome diagnostics of grain legumes. In: Kirti PB (ed) Handbook of new technologies for genetic improvement of legumes. CRC Press, New York, pp 497–557

    Google Scholar 

  • Sehgal D, Rajpal VR, Raina SN (2008c) Chloroplast DNA diversity reveals the contribution of two wild species to the origin and evolution of diploid Safflower (Carthamus tinctorius L.). Genome 51:638–643

    Article  CAS  PubMed  Google Scholar 

  • Sehgal D, Rajpal VR, Raina SN, Sasanuma T, Sasakuma T (2008d) Assaying polymorphism at DNA level for genetic diversity diagnostics of the safflower (Carthamus tinctorious L.) world germplasm resources. Genetica 135:457–470

    Article  PubMed  Google Scholar 

  • Shah A, De-Z Li, Moller M, Gao L-M, Hollingsworth ML, Gibby M (2008) Delimitation of Taxus fuana Nan Li & R.R. Mill (Taxaceae) based on morphological and molecular data. Taxon 57(1):211–222

    Google Scholar 

  • Shiponuv AB, Fay MF, Pillon Y, Bateman RM, Chase MW (2004) Dactylorhiza (Orchidaceae) in European Russia: combined molecular and morphological analysis. Am J Bot 91:1419–1426

    Article  Google Scholar 

  • Shiponuv AB, Fay MF, Pillon Y, Bateman RM, Chase MW (2005) The taxonomy of Dactylorhiza baltica (Orchidaceae) from European Russia: evidence from molecular markers and morphology. Bot J Linn Soc 147:257–274

    Article  Google Scholar 

  • Shiran B, Raina SN (2001) Evidence of rapid evolution and incipient speciation in Vicia sativa species complex based on nuclear and organellar RFLPs and PCR analysis. Genet Resour Crop Evol 48:519–532

    Article  Google Scholar 

  • Singh S, Kumar S (1998) Withania somnifera. The Indian Ginseng ashwagandha. Central Institute of Medicinal and Aromatic Plants, Lucknow

    Google Scholar 

  • Singh A, Devarumath RM, Rama Rao S, Singh VP, Raina SN (2008) Assessment of genetic diversity, phylogenetic relationships based on ribosomal DNA repeat unit length variation and internal transcribed spacer (ITS) sequences in chickpea (Cicer arietinum) cultivars and its wild species. Genet Resour Crop Evol 55:65–79

    Article  CAS  Google Scholar 

  • Stebbins GL (1950) Variation and evolution in plants. Columbia University Press, New York

    Google Scholar 

  • Thompson JD, Gibson TJ, Plewniak F, Mougin FJ, Higgins DG (1997) The Clustal X Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882

    Article  CAS  PubMed  Google Scholar 

  • Uma DP, Sharda AC, Emerson SF (1993) Antitumour and radiosensitizing effects of Withania somnifera (Ashwagandha) on transplantable mouse tumour, Sarcoma 180. Indian J Exp Biol 31:60–61

    Google Scholar 

  • Uma DP, Akagi K, Ostapenko V, Tanaka Y, Sugihara T (1996) Withaferin A: a new radiosensitizer from the Indian medicinal plant Withania somnifera. Int J Radiat Biol 69:193–197

    Article  Google Scholar 

  • White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenies. In: Innis MA, Gelfand DH, Shinsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic press, San Diego, pp 315–322

    Google Scholar 

Download references

Acknowledgments

We thank Dr. K. A. Suri, Head NPC Division, for helping in chemical analysis. Technical assistance by Sh. Madan Lal during field work is also acknowledged. Thanks are also due to the Council of Scientific and Industrial Research, Government of India, for financial support to Dr. D. Sehgal and Mr. T. H. Dar.

Author information

Authors and Affiliations

  1. Indian Institute of Integrative Medicine, Jammu, 180001, India

    Arun Kumar, Bilal A. Mir, Sushma Koul, Maharaj K. Kaul & Ghulam N. Qazi

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

    Deepmala Sehgal & Tanvir H. Dar

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

    Soom N. Raina

Authors
  1. Arun Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bilal A. Mir
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Deepmala Sehgal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tanvir H. Dar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sushma Koul
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Maharaj K. Kaul
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Soom N. Raina
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ghulam N. Qazi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Arun Kumar.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kumar, A., Mir, B.A., Sehgal, D. et al. Utility of a multidisciplinary approach for genome diagnostics of cultivated and wild germplasm resources of medicinal Withania somnifera, and the status of new species, W. ashwagandha, in the cultivated taxon. Plant Syst Evol 291, 141–151 (2011). https://doi.org/10.1007/s00606-010-0372-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00606-010-0372-4

Keywords

Search

Navigation