Skip to main content

Advertisement

SpringerLink
Log in

In Vitro Propagation and Phytochemical Assessment of Aconitum ferox Wall: A Threatened Medicinal Plant of Sikkim Himalaya

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

Abstract

This is the first report on in vitro propagation and phytochemical assessment of Aconitum ferox (Ranunculaceae), a threatened medicinal plant of Sikkim Himalaya. A simple and efficient in vitro propagation protocol through indirect shoot organogenesis has been established for A. ferox using root tip explants. Murashige and Skoog (MS) medium supplemented with 2.26 µM 2, 4-dichlorophenoxyacetic acid (2, 4-D) was found to be the best medium to induce and maintain the callus. Different concentrations and combinations of plant growth regulators (PGRs) were tested for in vitro shoot proliferation. Adventitious shoots were produced from callus when it was transferred to MS medium supplemented with N6-benzylaminopurine (BAP). The synergistic incorporation of 3 µM indole-3-acetic acid (IAA) in 6 µM BAP containing medium-induced plantlets with well-developed roots and shoots. However, the best rooting responses were observed in shoots placed on paper bridge in liquid MS medium supplemented with 3 µM IAA + 6 µM BAP. The plantlets were successfully acclimatized in ex vitro conditions with 70% survival rate. Additionally, root extract of in vitro-raised plants showed antioxidant activity closer to wild plants. The study thus signifies the effectiveness of in vitro technique for A. ferox propagation and provides a method for sustainable utilization of this high-value medicinal plant at commercial scale in pharmaceutical industries.

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

References

  1. Badola HK, Aitken S (2010) Potential biological resources for poverty alleviation in Indian Himalaya. Biodiversity 11(3–4):8–18. https://doi.org/10.1080/14888386.2010.9712659

    Article  Google Scholar 

  2. Ved DK, Kinhal GA, Haridasan K, Ravikumar K, Ghate U, Sankar RV, Indresha JH (2003) Conservation assessment and management prioritisation for the medicinal plants of Arunachal Pradesh. FRLHT, Bangalore, India, Assam, p 120

    Google Scholar 

  3. Pradhan BK, Badola HK (2015) Swertia chirayta, a threatened high-value medicinal herb: microhabitats and conservation challenges in Sikkim Himalaya, India. Mount Res Dev 35(4):374–381. https://doi.org/10.1659/MRD-JOURNAL-D-14-00034.1

    Article  Google Scholar 

  4. Nayar MP, Sastry APK (1990) Red data book of Indian plants. Botanical Survey of India, Calcutta

    Google Scholar 

  5. Chakravarty HL, Chakravarti D (1954) Indian aconites. Econ Bot 8:366–376

    Article  Google Scholar 

  6. Rawat JM, Agnihotri RK, Nautiyal S, Rawat B, Chandra A (2013) In vitro propagation, genetic and secondary metabolite analysis of Aconitum violaceum Jacq.: a threatened medicinal herb. Acta Physiol Plant 35:2589–2599. https://doi.org/10.1007/s11738-013-1294-x

    Article  CAS  Google Scholar 

  7. Uprety Y, Asselin H, Boon EK, Yadav S, Shrestha KK (2010) Indigenous use and bio-efficacy of medicinal plants in the Rasuwa district, central Nepal. J Ethnobiol Ethnomed 6(1):3. https://doi.org/10.1186/1746-4269-6-3

    Article  PubMed  PubMed Central  Google Scholar 

  8. Pradhan BK, Badola HK (2008) Ethnomedicinal plant use by Lepcha tribe of Dzongu valley, bordering Khangchendzonga Biosphere Reserve, in North Sikkim, India. J Ethnobiol Ethnomed 4:22. https://doi.org/10.1186/1746-4269-4-22

    Article  PubMed  PubMed Central  Google Scholar 

  9. Sheokand A, Sharma A, Gothecha VK (2012) Vatsanabha (Aconitum Ferox): from Visha to Amrita. Int J Ayurvedic Herb Med 2(3):423–426. https://doi.org/10.7897/2277-4343.085278

    Article  CAS  Google Scholar 

  10. Hanuman JB, Katz A (1994) New lipo norditerpenoid alkaloids from root tubers of Aconitum ferox. J Nat Prod 57:105–115. https://doi.org/10.1021/np50103a015

    Article  CAS  Google Scholar 

  11. Hanuman JB, Katz A (1994) Diterpenoid alkaloids from Ayurvedic processed and unprocessed Aconitum ferox. Phytochem 36:1527–1535. https://doi.org/10.1016/S0031-9422(00)89756-3

    Article  CAS  Google Scholar 

  12. Purushothman KK, Chandrasekharan S (1974) Alkaloids of Aconitum ferox. Phytochem 13:1975–1977. https://doi.org/10.1016/0031-9422(74)85128-9

    Article  Google Scholar 

  13. Bhatt D, Joshi GC, Kumar R, Tewari LM (2014) Phytosociological features and categorization of A. heterophyllum Wall. ex Royle and A. ferox Wall. ex Ser. in Kumaun Himalaya. J Ecol Nat Env 6(3):111–118. https://doi.org/10.5897/jene2013.0395

    Article  Google Scholar 

  14. Cervelli R (1987) In vitro propagation of Aconitum noveboracense and Aconitum napellus. Hort Sci 22(2):304–305

    CAS  Google Scholar 

  15. Hatano K, Kamura K, ShoyamaY NishiokaI (1988) Clonal multiplication of Aconitum carmichaeli Debx. by tip tissue culture and alkaloid contents of clonally propagated plants. Planta Med 54:152–154. https://doi.org/10.1055/s-2006-962375

    Article  CAS  PubMed  Google Scholar 

  16. Giri A, Ahuja PS, Kumar PVA (1993) Somatic embryogenesis and plant regeneration from callus culture of Aconitum heterophyllum Wall. Plant Cell Tiss Organ Cult 32:213–218. https://doi.org/10.1007/BF00029845

    Article  CAS  Google Scholar 

  17. Chandra B (2003) Studies on propagation, agrotechnology and phytochemical evaluation of some alpine medicinal plants of Himalayan region. Ph.D. thesis, Kumaun University, Nainital, p 148

  18. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco cultures. Physiologia Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

    Article  CAS  Google Scholar 

  19. Meda A, Lamien CE, Romito M, Millogo J, Nacoulma OG (2005) Determination of the total phenolic, flavonoid and proline contents in Burkina Fasan honey, as well as their radical scavenging activity. Food Chem 91(3):571–577. https://doi.org/10.1016/j.foodchem.2004.10.006

    Article  CAS  Google Scholar 

  20. Lamaison JLC, Carnet A (1990) Contents in main flavonoid compounds of Crataegus monogyna Jacq. and Crataegus laevigata (Poiret) DC flowers at different development stages. Pharmaceutica Acta Helvetica 65:315–320

    CAS  Google Scholar 

  21. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Evans C (1999) Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biol Med 26:1231–1237 PMID: 10381194

    Article  CAS  Google Scholar 

  22. Pandey H, Nandi SK, Kumar A, Palni UT, Chandra B, Palni LMS (2004) In vitro propagation of Aconitum balfourii Stapf; an important aconite of Himalayan alpine. J Hort Sci Biotech 21:69–84. https://doi.org/10.1080/14620316.2004.11511733

    Article  Google Scholar 

  23. Belwal NS, Kamal B, Sharma V, Gupta S, Dobriyal AK, Jadon VS (2016) Production of genetically uniform plant from shoot tips of Aconitum heterophyllum Wall.- a critically endangered medicinal herb. J Hortic Sci Biotechnol. https://doi.org/10.1080/14620316.2016.1184434

    Article  Google Scholar 

  24. Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp Soc Exp Biol 11:118–130 PMID:13486467

    CAS  PubMed  Google Scholar 

  25. Gordon SP, Chickarmane VS, Ohno C, Meyerowitz EM (2009) Multiple feedback loops through cytokinin signaling control stem cell number within the Arabidopsis shoot meristem. Proc Natl Acad Sci USA 106:16529–16534. https://doi.org/10.1073/pnas.0908122106

    Article  PubMed  Google Scholar 

  26. Singh M, Chaturvedi R (2012) Screening and quantification of an antiseptic alkylamide, spilanthol from in vitro cell and tissue cultures of Spilanthes acmella Murr. Ind Crop Prod 36:321–328. https://doi.org/10.1016/j.indcrop.2011.10.029

    Article  CAS  Google Scholar 

  27. Pandey A, Tamta S (2014) In vitro propagation of the important tasar Oak (Quercus serrata Thunb.) by casein hydrolysate promoted high frequency shoot proliferation. J Sust For 33:590–603. https://doi.org/10.1080/10549811.2014.912587

    Article  Google Scholar 

  28. Brijwal L, Pandey A, Tamta S (2015) In vitro propagation of the endangered species Berberis aristata DC. via leaf-derived callus. Vitro Cell Dev Biol- Plant 51:1–11. https://doi.org/10.1007/s11627-015-9716-7

    Article  CAS  Google Scholar 

  29. Hesami M, Daneshvar MH, Yoosefzadeh-Najafabadi M, Alizadeh M (2017) Effect of plant growth regulators on indirect shoot organogenesis of Ficus religiosa through seedling derived petiole segments. J Genet Eng Biotechnol 16:175–180. https://doi.org/10.1016/j.jgeb.2017.11.001

    Article  PubMed  PubMed Central  Google Scholar 

  30. Zhu WT, Du JZ, Sun HB, Jiang SY, Chen XJ, Sun H, Zhou Y, Wang HL (2018) In vitro propagation of an endemic and endangered medicinal plant Notopterygium incisum. Plant Cell Tiss Organ Cult. https://doi.org/10.1007/s11240-018-1474-x

    Article  Google Scholar 

  31. Pereira JA, Oliveira I, Sousa A, Valentão P, Andrade PB, Ferreira IC, Ferreres F, Bento A, Seabra R, Estevinho L (2007) Walnut (Juglans regia L.) leaves: phenolic compounds, antibacterial activity and antioxidant potential of different cultivars. Food Chem Toxicol 45(11):2287–2295. https://doi.org/10.1016/j.fct.2007.06.004

    Article  CAS  PubMed  Google Scholar 

  32. Covelo F, Gallardo A (2001) Temporal variation in total leaf phenolics concentration of Quercus robur in forested and harvested stands in Northwestern Spain. Canad J Bot 79(11):1262–1269. https://doi.org/10.1139/b01-109

    Article  CAS  Google Scholar 

  33. Munir N, Ijaz W, AltafI Naz S (2014) Evaluation of antifungal and antioxidant potential of two medicinal plants: Aconitum heterophyllum and Polygonum bistorta. Asian Pac J Tropical Biomed 4:639–643. https://doi.org/10.12980/APJTB.4.201414B182

    Article  Google Scholar 

  34. Rameashkannan MV, Riyaz AP, Mudasir AM (2013) Phytochemical, antioxidant and antimicrobial evaluation of Aconitum kashmiricum Stapf ex Coventry, a critically endangered medicinal herb. Indo Am J Pharm Res 4(1):2231–6876. https://doi.org/10.1044/1980-iajpr.01125

    Article  Google Scholar 

Download references

Acknowledgements

The authors are thankful to the Director, G.B. Pant National Institute of Himalayan Environment and Sustainable Development (GBPNIHESD), and to the Ministry of Environment Forest and Climate Change, (MoEF & CC), Govt. of India, New Delhi, for providing necessary facilities. Financial support from GBPNIHESD, Kosi-Katarmal, Almora, MoEF & CC (In House Project No. 8) is gratefully acknowledged. Mr. Mohan Thapa, GBPNIHESD, Sikkim Regional Centre is acknowledged for his help during experimentation.

Author information

Authors and Affiliations

  1. G.B. Pant, National Institute of Himalayan Environment and Sustainable Development, Sikkim Regional Centre, Pangthang, Gangtok, Sikkim, 737101, India

    Mithilesh Singh, Archana Chettri, Aseesh Pandey, Sheila Sinha & K. K. Singh

  2. D-408, Aditya Doonshire Apartments, Sailok Phase II, GMS Road, Dehradun, Uttarakhand, 248001, India

    H. K. Badola

Authors
  1. Mithilesh Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Archana Chettri
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Aseesh Pandey
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sheila Sinha
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. K. K. Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. H. K. Badola
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mithilesh Singh.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest to publish this manuscript.

Additional information

Publisher's Note

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

Significance Statement This is the first report on in vitro propagation and phytochemical assessment of A. ferox. Study provides a sustainable technique for germplasm conservation and commercial cultivation of this threatened and high-value medicinal plant of Himalaya.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Singh, M., Chettri, A., Pandey, A. et al. In Vitro Propagation and Phytochemical Assessment of Aconitum ferox Wall: A Threatened Medicinal Plant of Sikkim Himalaya. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 90, 313–321 (2020). https://doi.org/10.1007/s40011-019-01104-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40011-019-01104-x

Keywords

Search

Navigation