Skip to main content
Log in

Clonal fidelity and phytochemical analysis of in vitro propagated Kaempferia rotunda Linn.—an endangered medicinal plant

  • Medicinal Plants
  • Published:
In Vitro Cellular & Developmental Biology - Plant Aims and scope Submit manuscript

Abstract

Kaempferia rotunda Linn. (Zingiberaceae) is an important herb that has both therapeutic and cosmetic applications. An efficient protocol has been developed for in vitro propagation of K. rotunda using axillary bud explants from unsprouted rhizomes. Murashige and Skoog medium containing 3.0 mg L−1 6-Benzyladenine (BA) in combination with 1.0 mg L−1 indoleacetic acid (IAA) was found to be optimum for the regeneration, multiplication, and in vitro maintenance of plantlets. Two-yr-old ex vitro grown micropropagated plants were assessed for stable drug-yielding potential through the evaluation of essential oil contents, its phytoconstituents, and antioxidant activity. Gas chromatography and mass spectroscopy (GCMS) analysis of essential oil of rhizome showed the presence of benzoic acid (61.34% and 58.27%), bornyl ester (15.11% and 14.66%), zingiberene (5.15% and 5.74%), and camphor (3.72% and 3.82%) in both micropropagated and conventionally grown K. rotunda, respectively. Methanolic extracts and essential oils of the rhizome of both plants possess almost the same antioxidant activity as revealed from DPPH free radical scavenging assay. Micropropagated K. rotunda also proved to be genetically stable as revealed by RAPD and ISSR-based molecular profiling. Thus, this study concluded that micropropagation of Kaempferia rotunda, an endangered medicinal plant, can be recommended for large-scale commercial production of true-to-type plantlets with stable drug-yielding potential.

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

Access this article

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

Instant access to the full article PDF.

Figure 1.
Figure 2.
Figure 3.
Figure 4.

Similar content being viewed by others

References

  • Adams RP (2007) Identification of essential oils by gas chromatography/mass spectrometry. Allured Publishing Corporation, Carol Stream

    Google Scholar 

  • Agnihotri RK, Mishra J, Nandi SK (2009) Improved in vitro shoot multiplication and rooting of Dendrocalamus hamiltonii Nees et Arn. Ex Munro: production of genetically uniform plants and field evaluation. Acta Physiol Plant 31:961–967

    Article  CAS  Google Scholar 

  • Ajay K (2014) Chemical characterization of essential oil from the rhizomes of Kaempferia rotunda L. by GC/MS technique. Int J Pharm Biol Sci 5:458–462

    Google Scholar 

  • Amoo SO, Aremu AO, Van Staden J (2012) In vitro plant regeneration, secondary metabolite production and antioxidant activity of micropropagated Aloe arborescens Mill. Plant Cell Tiss Org Cult 111:345–358

    Article  CAS  Google Scholar 

  • Behera B, Sinha P, Gouda S, Rath SK, Barik DP, Jena PK, Panda PC, Naik SK (2018) In vitro propagation by axillary shoot proliferation, assessment of antioxidant activity, and genetic fidelity of micropropagated Paederia foetida L. J App Biol Biotech 6:41–49

    CAS  Google Scholar 

  • Behera S, Kamila PK, Rout KK, Barik DP, Panda PC, Naik SK (2018) An efficient plant regeneration protocol of an industrially important plant, Hedychium coronarium J. Koenig and establishment of genetic and biochemical fidelity of the regenerants. Ind Crops Prod 126:58–68

    Article  CAS  Google Scholar 

  • Behera S, Kar SK, Rout KK, Barik DP, Panda PC, Naik SK (2019) Assessment of genetic and biochemical fidelity of field-established Hedychium coronarium J. Koenig regenerated from axenic cotyledonary node on meta-topolin supplemented medium. Ind Crops Prod 134:206–215

    Article  CAS  Google Scholar 

  • Behera S, Monalisa K, Meher RK, Mohapatra S, Madkami SK, Das PK, Naik PK, Naik SK (2022) Phytochemical fidelity and therapeutic activity of micropropagated Curcuma amada Roxb.: a valuable medicinal herb. Ind Crops Prod 176:114401

    Article  CAS  Google Scholar 

  • Bejoy M, Dan M, Anish NP (2006) Factors affecting the in vitro multiplication of the endemic zingiber Curcuma haritha Mangaly and Sabu. Asian J Plant Sci 5:847–853

    Article  Google Scholar 

  • Chan LK, Thong WH (2004) In vitro propagation of Zingiberaceae species with medicinal properties. J Plant Biotechnol 6:181–188

    Google Scholar 

  • Chen SL, Yu H, Luo HM, Wu Q, Li CF, Steinmetz A (2016) Conservation and sustainable use of medicinal plants: problems, progress, and prospects. Chin Med 11:1–10

    Article  PubMed  PubMed Central  Google Scholar 

  • Chirangini P, Sinha SK, Sharma GJ (2005) In vitro propagation and microrhizome induction in Kaempferia galangal Linn. and Kaempheria rotunda Linn. Indian J Biotechnol 4:404–408

    Google Scholar 

  • Das Bhowmik SS, Basu A, Sahoo L (2016) Direct shoot organogenesis from rhizomes of medicinal Zingiber Alpinia calcarata Rosc. and evaluation of genetic stability by RAPD and ISSR markers. J Crop Sci Biotechnol 19:157–165

    Article  Google Scholar 

  • Doyle JJ, Doyle JL (1990) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15

    Google Scholar 

  • Feng AS (2009) Chemical Constituents and Bioactivity of Malaysian and Indonesian Kaempferia rotunda (Doctoral dissertation, M. Sc. Thesis, Faculty of Science, University of Teknologi, Malaysia)

  • Geetha SP, Manjula C, John CZ, Minoo D, Nirmal Babu K, Ravindran PN (1997) Micropropagation of Kaempferia spp. (K. galanga L. and K. rotunda L.). J Spices Aromat Crops 6:129–135

    Google Scholar 

  • Giri L, Jugran A, Rawat S, Dhyani P, Andola H, Bhatt ID, Rawal RS, Dhar U (2012) In vitro propagation, genetic and phytochemical assessment of Habenaria edgeworthii: an important Astavarga plant. Acta Physiol Plant 34:869–875

    Article  Google Scholar 

  • Jamalluddin NAC (2014) Essential oils, phytochemicals and bioactivity studies of Curcuma aeruginosa Aff. and Kaempferia rotunda Linn (Doctoral dissertation, Universiti Teknologi Malaysia)

  • Jena S, Ray A, Sahoo A, Sahoo S, Dash B, Kar B, Nayak S (2020) Rapid plant regeneration in industrially important Curcuma zedoaria revealing genetic and biochemical fidelity of the regenerants. 3 Biotech 10:1–13

    Article  Google Scholar 

  • Jena S, Ray A, Sahoo A, Sahoo S, Kar B, Panda PC, Nayak S (2018) High-frequency clonal propagation of Curcuma angustifolia ensuring genetic fidelity of micropropagated plants. Plant Cell Tiss Org Cult 135:473–486

    Article  CAS  Google Scholar 

  • Mohanty S, Joshi RK, Subudhi E, Sahoo S, Nayak S (2010) Assessment of genetic stability of micropropagated Curcuma caesia through cytophotometric and molecular analysis. Cytologia 75:73–81

    Article  CAS  Google Scholar 

  • Mohanty S, Panda MK, Subudhi E, Acharya L, Nayak S (2008) Genetic stability of micropropagated ginger derived from axillary bud through cytophotometric and RAPD analysis. Z Naturforsch 63:747–754

    Article  CAS  Google Scholar 

  • Mohanty S, Parida R, Singh S, Joshi RK, Subudhi E, Nayak S (2011) Biochemical and molecular profiling of micropropagated and conventionally grown Kaempferia galanga. Plant Cell Tiss Org Cult 106:39–46

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Mustafaanand PH (2014) In-vitro plant regeneration in Kaempferia rotunda Linn. through somatic embryogenesis-a rare medicinal plant. Int J Curr Microbiol Appl Sci 3:409–414

    Google Scholar 

  • Nair RV (2004) Controversial drug plants. Universities Press

    Google Scholar 

  • Nayak S (2002) High frequency in vitro production of microrhizomes of Curcuma amada. Indian J Exp Biol 40:230–232

    PubMed  Google Scholar 

  • Nayak S, Kaur T, Mohanty S, Ghosh G, Choudhury R, Acharya L, Subudhi E (2011) In vitro and ex vitro evaluation of long-term micropropagated turmeric as analyzed through cytophotometry, phytoconstituents, biochemical and molecular markers. Plant Growth Regul 64:91–98

    Article  CAS  Google Scholar 

  • Panda MK, Mohanty S, Subudhi E, Acharya L, Nayak S (2007) Assessment of genetic stability of micropropagated plants of Curcuma longa L. by cytophotometry and RAPD analysis. Int J Integr Biol 1:189–195

    CAS  Google Scholar 

  • Parida R, Mohanty S, Kuanar A, Nayak S (2010) Rapid multiplication and in vitro production of leaf biomass in Kaempferia galanga through tissue culture. Electron J Biotechnol 13:1–8

    Article  Google Scholar 

  • Pittampalli B, Jogam P, Thampu RK, Abbagani S, Peddaboina V (2022) High-frequency plant regeneration and genetic homogeneity assessment of regenerants by molecular markers in turmeric (Curcuma longa L.). In Vitro Cell Dev Biol - Plant 58:169–180

    Article  CAS  Google Scholar 

  • Purohit S, Nandi SK, Paul S, Tariq M, Palni LMS (2017) Micropropagation and genetic fidelity analysis in Amomum subulatum Roxb.: a commercially important Himalayan plant. J Appl Res Med Aromat Plants 4:21–26

    Google Scholar 

  • Rakkimuthu R, Jacob J, Aravinthan KM (2011) In vitro micropropagation of Alpinia zerumbet Variegate, an important medicinal plant, through rhizome bud explants. Res Biotechnol 2:07–10

    Google Scholar 

  • Rao CK, Geetha BL, Suresh G (2003) Red list of threatened vascular plant species in India. Botanical Survey of India, Kolkata

    Google Scholar 

  • Ray T, Dutta I, Saha P, Das S, Roy SC (2006) Genetic stability of three economically important micropropagated banana (Musaspp.) cultivars of lower Indo-Gangetic plains, as assessed by RAPD and ISSR markers. Plant Cell Tiss Org Cult 85:11–21

    Article  CAS  Google Scholar 

  • Sahoo S, Ghosh G, Das D, Nayak S (2013) Phytochemical investigation and in vitro antioxidant activity of an indigenous medicinal plant Alpinia nigra BL Burtt. Asian Pac J Trop Biomed 3:871–876

    Article  CAS  PubMed Central  Google Scholar 

  • Sahoo S, Singh S, Nayak S (2014) Chemical composition, antioxidant and antimicrobial activity of essential oil and extract of Alpinia malaccensis Roscoe (Zingiberaceae). Int J Pharm Pharm Sci 6:183–188

    CAS  Google Scholar 

  • Sahoo S, Singh S, Sahoo A, Sahoo BC, Jena S, Kar B, Nayak S (2020) Molecular and phytochemical stability of long term micropropagated greater galanga (Alpinia galanga) revealed suitable for industrial applications. Ind Crops Prod 148:112274

    Article  CAS  Google Scholar 

  • Sereena K, Prakash KU, Rema SAB (2011) Histochemical and phytochemical markers for the authentication of Ayurvedic raw drug Hallakam (Kaempferia rotunda) and its marketed adulterant. Int J Pharm Sci Res 2:2952–2958

    Google Scholar 

  • Singh S, Kuanar A, Mohanty S, Subudhi E, Nayak S (2011) Evaluation of phytomedicinal yield potential and molecular profiling of micropropagated and conventionally grown turmeric (Curcuma longa L.). Plant Cell Tiss Org Cult 104:263–269

    Article  CAS  Google Scholar 

  • Sirat HM, Jamil S, Siew LW (2005) The rhizome oil of Kaempferia rotunda Val. J Essent Oil Res 17:306–307

    Article  CAS  Google Scholar 

  • Tyagi RK, Agrawal A, Mahalakshmi C, Hussain Z, Tyagi H (2007) Low-cost media for in vitro conservation of turmeric (Curcuma longa L.) and genetic stability assessment using RAPD markers. In Vitro Cell Dev Biol - Plant 43:51–58

    Article  CAS  Google Scholar 

  • Udayan PS, Balachandran I (2009) Medicinal plants of Arya Vaidya Sala Herb Garden. Kottakkal Ayurveda Series: 72. Department of Publications, Arya Vaidya Sala, Kottakkal, Kerala

  • Venkatachalam L, Sreedhar RV, Bhagyalakshmi N (2007) Molecular analysis of genetic stability in long-term micropropagated shoots of banana using RAPD and ISSR markers. Electron J Biotechnol 15:106–113

    Google Scholar 

  • Virk PS, Zhu J, Newbury HJ, Bryan GJ, Jackson MT, Ford-Lloyd BV (2000) Effectiveness of different classes of molecular marker for classifying and revealing variation in rice (Oryza sativa) germplasm. Euphytica 112:275–284

    Article  CAS  Google Scholar 

  • Wink M (2015) Modes of action of herbal medicines and plant secondary metabolites. Medicines 2:251–286

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Woerdenbag HJ, Windono T, Bos R, Riswan S, Quax WJ (2004) Composition of the essential oils of Kaempferia rotunda L. and Kaempferia angustifolia Roscoe rhizomes from Indonesia. Flavour Fragr J 19:145–148

    Article  CAS  Google Scholar 

  • Xu S, Yin G, Liu J, Lu Y, Guo S, Shi H (2012) Analysis for volatile compounds of Kaempferia rotunda. J Yunnan Univ Nat Sci 34:701–704

    Google Scholar 

Download references

Acknowledgements

Authors are thankful to Dr. Sudam Chandra Si, Dean Centre for Biotechnology, and Dr. Manoj Ranjan Nayak, President, Siksha O Anusandhan (Deemed to be University) for their support and encouragement throughout.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Suprava Sahoo.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sahoo, S., Lenka, J., Kar, B. et al. Clonal fidelity and phytochemical analysis of in vitro propagated Kaempferia rotunda Linn.—an endangered medicinal plant. In Vitro Cell.Dev.Biol.-Plant 59, 329–339 (2023). https://doi.org/10.1007/s11627-023-10342-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11627-023-10342-8

Keywords

Navigation