Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 135, Issue 3, pp 499–513 | Cite as

In vitro propagation, clonal fidelity and phytochemical analysis of Rhodiola imbricata Edgew: a rare trans-Himalayan medicinal plant

  • Ashwani Kumar Bhardwaj
  • Baldev Singh
  • Kulwinder Kaur
  • Poonam Roshan
  • Anu Sharma
  • Dechen Dolker
  • Avilekh Naryal
  • Shweta Saxena
  • Pratap Kumar PatiEmail author
  • Om Prakash ChaurasiaEmail author
Original Article


The present study focuses on development of a micropropagation protocol for true to type plants of Rhodiola imbricata, an endangered medicinal plant found in trans-Himalayan Leh-Ladakh region of India. It also aims at analyzing the pharmaceutically important secondary metabolites and antioxidant potential of in vitro and in vivo plants. Various cytokinins and auxins were tested for shoot proliferation and in vitro rooting of the microshoots, respectively. Random primers were used for checking genetic uniformity at different stages of micropropagation. Pharmaceutically important secondary metabolites of R. imbricata such as Rosavin, total polyphenols and free radical scavenging activity were analyzed by HPLC. Among different cytokinins used, BAP (5 µM) and TDZ (1 µM) were found to perform better in terms of shoot proliferation, shoot length and number of leaves as compared to other concentrations. For rooting of microshoots, a lower concentration of NAA (0.5 µM) yielded more efficient rooting of micro shoots (17.33 roots per micro shoot). In vitro rooted microshoots were hardened and showed 60% survival rate. The content of gallic acid, chlorogenic acid and 4-hydroxybenzoic acid was higher in the in vivo plant. The amount of ferulic acid was higher in the in vitro raised plant when compared to field grown plant. Furthermore, caffeic acid and p-coumaric acid were higher in the in vitro raised plants as compared to field grown plants. This work will facilitate in conservation of this endangered herb and provide necessary plant materials for various biotechnological and pharmaceutical applications.


Rhodiola imbricata Conservation Micropropagation Clonal fidelity Antioxidant Rosavin 



Analysis of variance


6-Benzyl amino purine


Cetyl trimethyl ammonium bromide




Fluorescein diacetate


Gibberellic acid


High performance liquid chromatography


Indole-3-acetic acid


Indole-3-butyric acid




Naphthalene acetic acid


Random amplified polymorphic DNA




Total polyphenolic content


Triphenyl tetrazolium chloride



The authors are thankful to Defence Research and Development Organization (DRDO), Ministry of Defence, Government of India for providing financial assistance for the research.

Author contributions

AKB, PKP developed the micropropagation protocol. AKB, BS, KK, PR wrote the manuscript. AKB, BS, KK, PR, AS, DD and AN performed the experiments. PKP, SS, OPC supervised the experimental tasks and the writing of the manuscript. KK, BS, PR and PKP have done English editing.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Ashwani Kumar Bhardwaj
    • 1
  • Baldev Singh
    • 2
  • Kulwinder Kaur
    • 2
  • Poonam Roshan
    • 2
  • Anu Sharma
    • 2
  • Dechen Dolker
    • 2
  • Avilekh Naryal
    • 1
  • Shweta Saxena
    • 1
  • Pratap Kumar Pati
    • 2
    Email author
  • Om Prakash Chaurasia
    • 1
    Email author
  1. 1.Department of Biotechnology, Defence Institute of High Altitude ResearchDefence Research & Development OrganizationLehIndia
  2. 2.Department of BiotechnologyGuru Nanak Dev UniversityAmritsarIndia

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