Skip to main content
SpringerLink
Log in

Molecular characterization of India Ginseng Withania somnifera (L) using ISSR markers

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Background

Ashwagandha (Withania somnifera (L.) Dunal), popularly known as Indian ginseng or winter cherry is a multipurpose plant of immense therapeutic value in the ayurvedic and indigenous medicine system and distributed in wide geographic locations and exhibiting extensive phenotypic and chemical variability.

Methods and results

The present study was carried out to assess the molecular genetic diversity among 4 CIMAP varieties and five local cultivars of ashwagandha and cluster dendrograms were created by using 20 ISSR primers. A total of 224 bands of varied length were produced, out of which 193 (86.1%) products were polymorphic and 31 (13.8%) products were monomorphic. Where each ISSR arbitrary primer had 5–16 valuable bands with an average of 11.2 bands per primer, of which 86.16% bands were polymorphic. The PIC values ranged from 0.16 to 0.36 with an average PIC value of 0.29 and RP values ranged from 2.22 to 7.99. The UPGMA cluster analysis of 20 ISSR primers grouped the nine accessions into 2 major clusters. The first and second major cluster consists of seven and two accessions respectively.

Conclusion

Therefore, this study provides evidence that ISSR based molecular diversity assessment can be used as an efficient tool for detecting similarity and phylogenetic relationships among genotypes of Withania somnifera collected from different geographical locations. This information can be used to improve root and other characteristics of ashwagandha genotypes and there is also scope for the development of high-yielding varieties by selecting diverse parents for crossing (based on the molecular diversity) from the present accessions.

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.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

Data availability

Not applicable.

Code availability

Not applicable.

Abbreviations

AFLP:

Amplified Fragment Length Polymorphism

CTAB:

Cetyl trimethyl Ammonium Bromide

ISSR:

Inter Simple Sequence Repeat

PCR:

Polymerase Chain Reaction

PIC:

Polymorphism Information Content

PPB:

Polymorphic Bands

Rp:

Resolving Power

SSR:

Simple Sequence Repeat

UPGMA:

Unweighted Pair Group Method with Arithmetic mean

References

  1. Chopra RN (1994) Glossary of Indian Medicinal Plants. Academic Publishers India, New Delhi

    Google Scholar 

  2. Devi PU, Sharada AC, Solomon FE (1993) Antitumor and radio sensitizing effects of Withania somnifera (Ashwagandha) on a transplantable mouse tumor, Sarcoma-180. Indian J Exp Biol. 31:607–11

    CAS  PubMed  Google Scholar 

  3. Singh S, Kumar S (1998) Withania somnifera: The Indian ginseng Ashwagandha. Central Institute of Medicinal and Aromatic Plants, Lucknow (India Stony Brook, New York, NY.)

    Google Scholar 

  4. Srivastava A, Anil KG, Shanker K, Madan MG, Mishra R, Raj KL (2018) Genetic variability, associations, and path analysis of chemical and morphological traits in Indian ginseng [Withania somnifera (L.) Dunal] for selection of higher yielding genotypes. J Ginseng Res 42:158–164. https://doi.org/10.1016/j.jgr.2017.01.014

    Article  PubMed  Google Scholar 

  5. Gupta AK, Verma SR, Gupta MM, Saikia D, Verma RK, Jhang T (2011) Genetic diversity in germplasms collections of Withania somnifera for root and leaf alkaloids. J Trop Med Plants 12:59–69

    Google Scholar 

  6. Al-Anbari AK, Al-Zubadiy MW, Malik Dawood W (2015) Genetic diversity of some taxa of Cucurbitaceae family based on RAPD markers. Adv Life Sci Technol 73:7–11

    Google Scholar 

  7. Yang HB, Kang WH, Nahm SH, Kang BC (2015) Methods for Developing Molecular Markers. In: Koh Hee-Jong, Kwon Suk-Yoon, Thomson Michael (eds) Current Technologies in Plant Molecular Breeding. Springer Science+Business Media, Dordrecht, pp 15–50

    Chapter  Google Scholar 

  8. Barvaliya P, Singh NK, Dharajiya D, Pachchigar K (2018) Elucidation of genetic diversity among ashwagandha [Withania somnifera (L.) Dunal] genotypes using EST-SSR markers. Res. J. Biotech. 13(10):52–59

    Google Scholar 

  9. Henareh MB, Abdollahi HK (2016) Assessment of genetic diversity in tomato landraces using ISSR markers. Genetika 48(1):25–35

    Article  Google Scholar 

  10. Ebrahimi M, Farajpour M, Rahimmalek M (2012) Inter- and intra-specific genetic diversity of Iranian yarrow species Achillea santolina and Achillea tenuifolia based on ISSR and RAPD markers. Genet Mol Res. https://doi.org/10.4238/2012.August.27.1

    Article  PubMed  Google Scholar 

  11. Farajpour M, Ebrahimi M, Amiri R, Golzari R, Sanjari S (2012) Assessment of genetic diversity in Achillea millefolium accessions from Iran using ISSR marker. Biochem Syst Ecol 43:73–79. https://doi.org/10.1016/j.bse.2012.02.017

    Article  CAS  Google Scholar 

  12. Maleki M, Shojaeiyan A, Monfared SR (2018) Population structure, morphological and genetic diversity within and among melon (Cucumis melo L.) landraces in Iran. J Genet Eng Biotechnol 16:599–606

    Article  Google Scholar 

  13. Baruah J, Pandey SK, Begum T, Sarma N, Paw M, Lal M (2019) Molecular diversity assessed amongst high dry rhizome recovery ginger germplasm (Zingiber officinale roscoe) from NE-India using RAPD and ISSR markers. Ind Crop Prod 129:463–471. https://doi.org/10.1016/j.indcrop.2018.12.037

    Article  CAS  Google Scholar 

  14. Saxena A (2014) Vol TR-RJ of B, 2020 U. assessment of genetic diversity in cowpea (Vigna unguiculata L. Walp.) germplasm. Bull Inst Trop Agric Kyushu Univ 37:57–63. https://doi.org/10.11189/bita.37.57

    Article  Google Scholar 

  15. Ghaffar K, Mohammad S (2017) Genetic Diversity in Tomato Varieties Assessed by ISSR Markers. International Journal of Vegetable Science 24(4):353–360. https://doi.org/10.1080/19315260.2017.1419397

    Article  Google Scholar 

  16. Farajpour M, Ebrahimi M, Amiri R, Noori SAS, Sanjari S, Golzari R (2011) Study of genetic variation in yarrow using inter-simple sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD) markers. Afr J Biotech 10:73–86

    Google Scholar 

  17. Khanuja SPS, Shasany AK, Darokar MP, Kumar S (1999) Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils. Plant Mol Biol Report 17(1):1–7

    Article  Google Scholar 

  18. Roldan RJ, Dendauw E, Van B, Depicker A, De Loose M (2000) AFLP markers reveal high polymorphic rates in ryegrasses (Lolium spp.). Mol Breed 6:125–134

    Article  Google Scholar 

  19. Prevost A, Wilkinson M (1999) A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor Appl Genet 98:107. https://doi.org/10.1007/s001220051046

    Article  CAS  Google Scholar 

  20. Rohlf FJ (1992) NTSYS-pc: Numerical taxonomy and multivariate analysis system. ver. 1.70.

  21. Pan L, Fu J, Zhang R et al (2017) Genetic diversity among germplasms of Pitaya based on SSR markers. Scientia Horticulturae 225:171–176

    Article  Google Scholar 

  22. Wolt JD (2019) Current risk assessment approaches for environmental and food and feed safety assessment. Transgenic Res 28:111–117. https://doi.org/10.1007/s11248-019-00140-7

    Article  CAS  PubMed  Google Scholar 

  23. Bilal AM, Sushma K, Kumar Arun, Kaul MK, Sooda AS, Raina SN (2009) Assessment and characterization of genetic diversity in Withania somnifera (L.) Dunal using RAPD and AFLP markers. African Journal of Biotechnology 10(66):14746–14756

    Google Scholar 

  24. Patel HK, Fougat RS, Kumar S, Mistry JG, Kumar M (2015) Detection of genetic variation in Ocimum species using RAPD and ISSR markers. 3 Biotech 5:697–707

    Article  Google Scholar 

  25. Gelotar MJ, Dharajiya DT, Solanki SD, Prajapati NN, Kapil KT (2019) Genetic diversity analysis and molecular characterization of grain amaranth genotypes using inter simple sequence repeat (ISSR) markers. Bull Natl Res Cent. https://doi.org/10.1186/s42269-019-0146-2

    Article  Google Scholar 

  26. Liu X, Jun D, Asaduzzaman Khan MD, Jingliang C, Chunli W, Zhiqiang M, Hanchun C, Tao H, Junjiang F (2020) Analysis of genetic diversity and similarities between different Lycium varieties based on ISSR analysis and RAMP-PCR markers. World Academy of Sci J 2:83–90

    Google Scholar 

  27. Gilberto CL, Danuza KS, Claudete AM, Maria de Fátima PSM (2020) ISSR markers to assess genetic diversity of cultivated populations from artificial selection of Stevia rebaudiana (Bert.) Bertoni. Breed Sci 70:508–514. https://doi.org/10.1270/jsbbs

    Article  Google Scholar 

  28. Khodaee L, Azizinezhad R, Etminan AR, Khosroshahi Mahmoud (2021) Assessment of genetic diversity among Iranian Aegilops triuncialis accessions using ISSR, SCoT, and CBDP markers. J Genet Eng Biotechnol. https://doi.org/10.1186/s43141-020-00107-w

    Article  PubMed  PubMed Central  Google Scholar 

  29. Niraj T, Navinder S, Vandana M, Sunil Kumar ST (2012) Assessment of genetic diversity among Withania somnifera collected from central India using RAPD and ISSR analysis. Medicinal and aromatic plant science and biotechnology 6(1):33–39

    Google Scholar 

  30. Žiarovská J, Ražná V, Labajová M (2013) Using of inter microsatellite polymorphism to evaluate gamma-irradiated Amaranth mutants. Emir J Food Agric 25(9):673–681

    Article  Google Scholar 

  31. Pfeiffer T, Roschanski AM, Pannell Korbecka G, Schnitter M (2011) Characterization of microsatellite loci and reliable genotyping in a polyploidy plant, Mercurialis perennis (Euphorbiaceae). J. Hered. 102:479–488

    Article  CAS  Google Scholar 

  32. Dharmar K, De John B (2011) RAPD analysis in genetic variability in wild populations of Withania somnifera (L.) Dunal. International journal of biological technology 2:21–25

    CAS  Google Scholar 

  33. Botstein D, White RL, Skolnick M, Davis RW (1980) Construction of a genetic linkage map in man using restriction fragment length polymorphisms. Am J Hum Genet 32:314–331

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Tiwari SK, Karihaloo JL, Hameed N, Ambika BG (2009) Molecular Characterization of Brinjal (Solanum melongena L) Cultivars using RAPD and ISSR Markers. J Plant Biochem Biotechnol 18:189–195. https://doi.org/10.1007/BF03263318

    Article  CAS  Google Scholar 

  35. Shahana K, Rehana AS (2016) Assessment of genetic diversity among India Ginseng, Withania somnifera (L) Dunal using RAPD and ISSR markers. Research in Biotechnology 7:01–10

    Google Scholar 

  36. Nayak S, Naik PK, Acharya L, Mukherjee AK, Panda PC, Das P (2005) Assessment of genetic diversity among 16 promising cultivars of ginger using cytological and molecular markers. Z Natur for schung 60:485–492

    CAS  Google Scholar 

  37. Kanika B, Sunita K, Anita P, Om Prakash Y (2013) Genetic Variability Analysis Using ISSR Markers in Withania Somnifera L. Dunal Genotypes from Different Regions. J Herbs Spices Med Plants 19(1):22–32. https://doi.org/10.1080/10496475.2012.734770

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors acknowledge the Director CSIR-CIMAP for providing the facility for conducting research work.

Funding

No Founding sources.

Author information

Authors and Affiliations

  1. CSIR—Central Institute of Medicinal and Aromatic Plants Research Centre, Bangalore, India

    Channayya Hiremath, Roja Philip & Velusamy Sundaresan

Authors
  1. Channayya Hiremath
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Roja Philip
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Velusamy Sundaresan
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

CH conceived the research idea and designed the experiments, and performed, analyzed the data, and wrote the manuscript. RP performed the experiments VS gave technical guidance while writing the manuscript and gave lab consumables for doing work.

Corresponding author

Correspondence to Channayya Hiremath.

Ethics declarations

Conflict of interest

The authors declare that there are no conflicts of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

CIMAP Publication No. CIMAP/PUB/2020/FEB /09.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hiremath, C., Philip, R. & Sundaresan, V. Molecular characterization of India Ginseng Withania somnifera (L) using ISSR markers. Mol Biol Rep 48, 3971–3977 (2021). https://doi.org/10.1007/s11033-021-06397-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-021-06397-8

Keywords

Search

Navigation